Efficient and quantitative high-throughput tRNA sequencing.

Abstract:

:Despite its biological importance, tRNA has not been adequately sequenced by standard methods because of its abundant post-transcriptional modifications and stable structure, which interfere with cDNA synthesis. We achieved efficient and quantitative tRNA sequencing in HEK293T cells by using engineered demethylases to remove base methylations and a highly processive thermostable group II intron reverse transcriptase to overcome these obstacles. Our method, DM-tRNA-seq, should be applicable to investigations of tRNA in all organisms.

journal_name

Nat Methods

journal_title

Nature methods

authors

Zheng G,Qin Y,Clark WC,Dai Q,Yi C,He C,Lambowitz AM,Pan T

doi

10.1038/nmeth.3478

subject

Has Abstract

pub_date

2015-09-01 00:00:00

pages

835-837

issue

9

eissn

1548-7091

issn

1548-7105

journal_volume

12

pub_type

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