Efficient and quantitative high-throughput tRNA sequencing.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 26214130.
- Also identified by DOI 10.1038/nmeth.3478 and PMC identifier 4624326.
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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.
Medical subject headings
- Algorithms
- Gene Library
- High-Throughput Nucleotide Sequencing
- RNA, Transfer