T-RHEX-RNAseq - a tagmentation-based, rRNA blocked, random hexamer primed RNAseq method for generating stranded RNAseq libraries directly from very low numbers of lysed cells

BMC Genomics. 2023 Apr 17;24(1):205. doi: 10.1186/s12864-023-09279-4.

Abstract

Background: RNA sequencing has become the mainstay for studies of gene expression. Still, analysis of rare cells with random hexamer priming - to allow analysis of a broader range of transcripts - remains challenging.

Results: We here describe a tagmentation-based, rRNA blocked, random hexamer primed RNAseq approach (T-RHEX-RNAseq) for generating stranded RNAseq libraries from very low numbers of FACS sorted cells without RNA purification steps.

Conclusion: T-RHEX-RNAseq provides an easy-to-use, time efficient and automation compatible method for generating stranded RNAseq libraries from rare cells.

Keywords: Expression profiling; RNA purification free; Random hexamer priming; Stranded RNAseq; Tagmentation.

MeSH terms

  • Base Sequence
  • Gene Expression Profiling / methods
  • High-Throughput Nucleotide Sequencing* / methods
  • RNA, Ribosomal* / genetics
  • Sequence Analysis, RNA / methods

Substances

  • RNA, Ribosomal