PRJDB16740 Development and application of TSS-seq2, a method for detecting transcription start sites with high specificity. We develop TSS-seq2, a method for detecting transcription start sites (TSS) with high specifity. To increase rate of vaild reads in 50 and 5 ng input libraries, we performed lower size-selection. To verfiy effect of long fragments within TSS-seq2 libraries, we also performed upper size removal of TSS-seq2 libraries prepared from 500 ng of total RNA. To demonstrate high specificity, TSS-seq2 libraries were prepared from 500 ng of degradated RNA whose DV200 values were 82% and 65%. For evaluation of quantitative measurements, we prepared TSS-seq2 libraries using 5 ng of ERCC Spike-in Mix. Development and application of TSS-seq2, a method for detecting transcription start sites with high specificity. bioproject PRJDB16740 PRJDB16740 true We develop TSS-seq2, a method for detecting transcription start sites (TSS) with high specifity. To increase rate of vaild reads in 50 and 5 ng input libraries, we performed lower size-selection. To verfiy effect of long fragments within TSS-seq2 libraries, we also performed upper size removal of TSS-seq2 libraries prepared from 500 ng of total RNA. To demonstrate high specificity, TSS-seq2 libraries were prepared from 500 ng of degradated RNA whose DV200 values were 82% and 65%. For evaluation of quantitative measurements, we prepared TSS-seq2 libraries using 5 ng of ERCC Spike-in Mix.