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Identification of nucleoside monophosphates and their epigenetic modifications using an engineered nanopore.

Nat Nanotechnol. 2022-07; 
Yuqin Wang , Shanyu Zhang , Wendong Jia ,Pingping Fan , Liying Wang , Xinyue Li , Jialu Chen , Zhenyuan Cao , Xiaoyu Du , Yao Liu , Kefan Wang , Chengzhen Hu , Jinyue Zhang , Jun Hu , Panke Zhang , Hong-Yuan Chen , Shuo Huang
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Gene Synthesis Te genes coding for monomeric M2 MspA-D16H6 (D90N/D91N/D93N/D118R/D134R/E139K) and N90C MspA-H6 (D90C/D91N/D93N/D118R/D134R/E139K) were separately synthesized and simultaneously inserted into a pET 30a(+) plasmid (GenScript). Get A Quote

摘要

RNA modifications play critical roles in the regulation of various biological processes and are associated with many human diseases. Direct identification of RNA modifications by sequencing remains challenging, however. Nanopore sequencing is promising, but the current strategy is complicated by sequence decoding. Sequential nanopore identification of enzymatically cleaved nucleoside monophosphates may simultaneously provide accurate sequence and modification information. Here we show a phenylboronic acid-modified hetero-octameric Mycobacterium smegmatis porin A nanopore, with which direct distinguishing between monophosphates of canonical nucleosides, 5-methylcytidine, N6-methyladenosine, N7-methylguanosine, N... More

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