Cost and time-efficient construction of a 3'-end mRNA library from unpurified bulk RNA in a single tube.
Exp Mol Med
; 56(2): 453-460, 2024 Feb.
Article
in En
| MEDLINE
| ID: mdl-38413820
ABSTRACT
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.
Full text:
1
Collection:
01-internacional
Health context:
1_ASSA2030
Database:
MEDLINE
Main subject:
RNA
/
Gene Expression Profiling
Language:
En
Journal:
Exp Mol Med
Year:
2024
Document type:
Article