DeLTa-Seq: direct-lysate targeted RNA-Seq from crude tissue lysate.

ABSTRACT:

Background

Quantification of gene expression such as RNA-Seq is a popular approach to study various biological phenomena. Despite the development of RNA-Seq library preparation methods and sequencing platforms in the last decade, RNA extraction remains the most laborious and costly step in RNA-Seq of tissue samples of various organisms. Thus, it is still difficult to examine gene expression in thousands of samples.

Results

Here, we developed Direct-RT buffer in which homogenization of tissue samples and direct-lysate reverse transcription can be conducted without RNA purification. The DTT concentration in Direct-RT buffer prevented RNA degradation but not RT in the lysates of several plant tissues, yeast, and zebrafish larvae. Direct reverse transcription on these lysates in

SUBMITTER: Kashima M 

PROVIDER: S-EPMC9356424 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature