3’Pool-seq: an optimized cost-efficient and scalable method of whole-transcriptome gene expression profiling (Run0064)

The advent of Next Generation Sequencing has allowed transcriptomes to be profiled with unprecedented accuracy, but the steep costs associated with full-length library sequencing have posed a limit on the accessibility and scalability of the technology. To address these limitations, we developed 3’Pool-seq, a simple, cost-effective, and scalable RNA-seq method that focuses sequencing to the 3’ end of mRNA transcripts. Transcriptomes (4 replicates each) from homozygous GFAP-IL6 mice were compared to those from wild-type C57BL/6 for differential gene expression using different amounts of cDNA input during the tagmentation step of 3'Pool-seq. In this specific experiment, different amounts of total RNA were added to the Reverse Transcription reaction in order to optimize that step for 3'Pool-seq.