Efficient and quantitative high-throughput tRNA sequencing
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 …
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 …
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.
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