SRX5893385 SBH17_S19 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 SBH17_S19 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893386 SBH16_S18 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 SBH16_S18 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893387 SBH15_S17 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 SBH15_S17 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893388 SBH14_S16 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 SBH14_S16 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893389 SBH10_S12 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 SBH10_S12 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893390 SBH06_S8 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 SBH06_S8 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893391 SBH05_S7 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 SBH05_S7 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893392 SBH04_S6 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 SBH04_S6 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893393 G416-TSN01 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G416-TSN01 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893394 SBH18_S20 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 SBH18_S20 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893395 G389-TSN17 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G389-TSN17 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893396 G338-TSN1 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G338-TSN1 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893397 G389-TSN14 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G389-TSN14 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893398 G389-TSN16 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G389-TSN16 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893399 G389-TSN11 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G389-TSN11 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893400 G389-TSN13 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G389-TSN13 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893401 G416-TSN17 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G416-TSN17 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893402 G389-TSN10 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G389-TSN10 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893403 G338-TSN5 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G338-TSN5 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893404 G338-TSN2 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G338-TSN2 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893405 G338-TSN3 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G338-TSN3 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893406 G338-TSN6 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G338-TSN6 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893407 G338-TSN4 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G338-TSN4 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893408 G416-TSN07 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G416-TSN07 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893409 G416-TSN05 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G416-TSN05 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893410 G416-TSN04 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G416-TSN04 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893411 G416-TSN02 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G416-TSN02 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893412 G416-TSN13 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G416-TSN13 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893413 G416-TSN11 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G416-TSN11 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893414 G416-TSN10 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G416-TSN10 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893415 G416-TSN08 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G416-TSN08 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893416 G416-TSN16 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G416-TSN16 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500 SRX5893417 G416-TSN14 SRP199445 NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, cells were collected and their genomic DNA (gDNA) was isolated as detailed above. Sequencing libraries were prepared using primers listed in Supplementary Table 4. In the first PCR step, the targeted gene was amplified from 100 ng of gDNA in a 25 l reaction with Q5 Master Mix (M0494L, NEB) and 500 nM final concentration of forward and reverse primers. This step was omitted for experiments in which the FANCM and SPRTN genes were targeted, as they did not include delivery of donor molecules. The thermal cycler program for the first PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 66C, 30 s at 72C, and 2 min at 72C. The second PCR was performed to add the Illumina P5 and P7 adaptors. The product from the first PCR was diluted at 100X and 2 l of this dilution was used as template. The thermal cycler program for the second PCR was as follows: 1 min at 98C, 35 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. A third PCR was performed to add index barcodes to each sample. For this PCR, the product of the first PCR was diluted 100X and 8 l of this dilution were used as template in a 25 l reaction with Q5 Master Mix and 500 nM final concentration of each of the forward (i5) and reverse (i7) primers. The thermal cycler program for the third PCR was as follows: 1 min at 98C, 12 cycles 10 s at 98C, 20 s at 60C, 30 s at 72C, and 2 min at 72C. PCR amplifications were verified using 2% agarose gels in TAE. The indexed amplicons from PCR #3 were pooled and gel purified. Gel purified samples were sequenced at the Genome Sciences Facility at The Pennsylvania State College of Medicine. SRS4815026 G416-TSN14 AMPLICON GENOMIC PCR 200 0 Application Read Forward 1 1 Application Read Reverse 101 Illumina HiSeq 2500