CAS NO: | 1612756-29-2 |
规格: | 98% |
分子量: | 228.21 |
包装 | 价格(元) |
5mg | 电议 |
10mg | 电议 |
25mg | 电议 |
50mg | 电议 |
100mg | 电议 |
Background:
NAI-N3 is a novel RNA acylation reagent that enables RNA purification.
The bifunctional NAI-N3 SHAPE reagent is used to label all RNAs inside living cells (or in vitro refolded RNA) to generate covalent modifications on targeted RNAs at single-nucleotide resolution. RNA is isolated, biotinylated using click chemistry and randomly fragmented to a size of 50-100 nt. To generate cDNA sequencing libraries, the RNA fragments are subjected to 3′-end repair, adapter ligation, size selection, biotin enrichment, reverse transcription (RT) and PCR. The end result is selective amplification of fragments corresponding to modified bases in the NAI-N3-treated samples or background expression values from mock-treated samples. Sequencing reads generated from the NAI-N3- and mock (DMSO)-treated samples are mapped to a transcriptome annotation, followed by computational identification of RT stops corresponding to SHAPE modifications—and hence reactive nucleotides—across the entire transcriptome[1].
[1]. Flynn RA, et al. Transcriptome-wide interrogation of RNA secondary structure in living cells with icSHAPE. Nat Protoc. 2016 Feb;11(2):273-90.