SRX7292768 ST SRP235324 bp0 RNA sequencing libraries were prepared with Illumina-compatible SureSelect Strand-SpecificRNA Library Prep kit (Agilent, Santa Clara, CA, U.S.A.) at the Genotypic Technology Pvt. Ltd., Bangalore,India.Briefly, mRNA enrichment was performed from 500ng of Qubit quantified total RNA. Theenriched mRNA was fragmented for 4 minutes at 94oC in the presence of divalent cations. First strandcDNA was synthesized in the presence of Actinomycin D (Gibco, life technologies, Carlsbad, CA, U.S.A.)and subsequently the single stranded cDNA was purified using HighPrep magnetic beads (MagbioGenomics Inc, USA). Purified single stranded cDNA was used for second strand synthesis followed byend repair using Second Strand Synthesis + End Repair mix. The end repaired cDNA was purified usingHighPrep magnetic beads. Adapters were ligated to the cDNA molecules after 3 Adenylation.The adapters used in the study were Illumina Universal Adapter:5-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT-3 andIndex Adapter:5-GATCGGAAGAGCACACGTCTGAACTCCAGTCAC[INDEX]ATCTCGTATGCCGTCTTCTGCTTG-3.[INDEX] Unique sequence to identify sample-specific sequencing data (Table 1)The adapter-ligated DNA was purified with HighPrep beads and then amplified for 10 cycles ofPCR using Illumina-compatible indexing primers provided in the SureSelect Strand-Specific RNA LibraryPrep kit. The final PCR product (sequencing library) was purified with HighPrep beads, followed bylibrary quality control check. The sequencing library was initially quantified by Qubit fluorometer(Thermo Fisher Scientific, MA, USA) and its fragment size distribution was analyzed on AgilentTapeStation (Table 1 and Appendix). Finally, the sequencing library was accurately quantified byquantitative PCR using Kapa Library Quantification Kit (Kapa Biosystems, Wilmington, MA, USA).The qPCR quantified libraries were pooled in equimolar amounts to create a final multiplexedlibrary pool for sequencing on an Illumina sequencer for 75 bp paired-end chemistry. SRS5786278 E.tirucalli P.ST ST RNA-Seq TRANSCRIPTOMIC Oligo-dT NextSeq 500 SRX7292769 SB SRP235324 bp0 RNA sequencing libraries were prepared with Illumina-compatible SureSelect Strand-SpecificRNA Library Prep kit (Agilent, Santa Clara, CA, U.S.A.) at the Genotypic Technology Pvt. Ltd., Bangalore,India.Briefly, mRNA enrichment was performed from 500ng of Qubit quantified total RNA. Theenriched mRNA was fragmented for 4 minutes at 94oC in the presence of divalent cations. First strandcDNA was synthesized in the presence of Actinomycin D (Gibco, life technologies, Carlsbad, CA, U.S.A.)and subsequently the single stranded cDNA was purified using HighPrep magnetic beads (MagbioGenomics Inc, USA). Purified single stranded cDNA was used for second strand synthesis followed byend repair using Second Strand Synthesis + End Repair mix. The end repaired cDNA was purified usingHighPrep magnetic beads. Adapters were ligated to the cDNA molecules after 3 Adenylation.The adapters used in the study were Illumina Universal Adapter:5-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT-3 andIndex Adapter:5-GATCGGAAGAGCACACGTCTGAACTCCAGTCAC[INDEX]ATCTCGTATGCCGTCTTCTGCTTG-3.[INDEX] Unique sequence to identify sample-specific sequencing data (Table 1)The adapter-ligated DNA was purified with HighPrep beads and then amplified for 10 cycles ofPCR using Illumina-compatible indexing primers provided in the SureSelect Strand-Specific RNA LibraryPrep kit. The final PCR product (sequencing library) was purified with HighPrep beads, followed bylibrary quality control check. The sequencing library was initially quantified by Qubit fluorometer(Thermo Fisher Scientific, MA, USA) and its fragment size distribution was analyzed on AgilentTapeStation (Table 1 and Appendix). Finally, the sequencing library was accurately quantified byquantitative PCR using Kapa Library Quantification Kit (Kapa Biosystems, Wilmington, MA, USA).The qPCR quantified libraries were pooled in equimolar amounts to create a final multiplexedlibrary pool for sequencing on an Illumina sequencer for 75 bp paired-end chemistry. SRS5786279 E.tirucalli P.SB SB RNA-Seq TRANSCRIPTOMIC Oligo-dT NextSeq 500