3'Pool-seq: an optimized cost-efficient and scalable method of whole-transcriptome gene expression profiling.

ABSTRACT:

Background

The advent of Next Generation Sequencing has allowed transcriptomes to be profiled with unprecedented accuracy, but the high costs of full-length mRNA sequencing have posed a limit on the accessibility and scalability of the technology. To address this, we developed 3'Pool-seq: a simple, cost-effective, and scalable RNA-seq method that focuses sequencing to the 3'-end of mRNA. We drew from aspects of SMART-seq, Drop-seq, and TruSeq to implement an easy workflow, and optimized parameters such as input RNA concentrations, tagmentation conditions, and read depth specifically for bulk-RNA.

Results

Thorough optimization resulted in a protocol that takes less than 12 h to perform, does not require custom sequencing primers or instrumentation, and cuts over 90% of the co

SUBMITTER: Sholder G 

PROVIDER: S-EPMC6971924 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature