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Cost and time-efficient construction of a 3'-end mRNA library from unpurified bulk RNA in a single tube.

Jungwon ChoiJungheun HyunJieun HyunJae-Hee KimJi Hyun LeeDuhee Bang
Published in: Experimental & molecular medicine (2024)
The major drawbacks of RNA sequencing (RNA-seq), a remarkably accurate transcriptome profiling method, is its high cost and poor scalability. Here, we report a highly scalable and cost-effective method for transcriptomics profiling called Bulk transcriptOme profiling of cell Lysate in a single poT (BOLT-seq), which is performed using unpurified bulk 3'-end mRNA in crude cell lysates. During BOLT-seq, RNA/DNA hybrids are directly subjected to tagmentation, and second-strand cDNA synthesis and RNA purification are omitted, allowing libraries to be constructed in 2 h of hands-on time. BOLT-seq was successfully used to cluster small molecule drugs based on their mechanisms of action and intended targets. BOLT-seq competes effectively with alternative library construction and transcriptome profiling methods.
Keyphrases
  • single cell
  • rna seq
  • small molecule
  • nucleic acid
  • high resolution
  • binding protein
  • cell free
  • mass spectrometry
  • wastewater treatment
  • protein protein
  • genome wide
  • mesenchymal stem cells
  • circulating tumor