Analysis of RNA-TAG labeling in HeLa lysates. Analysis of RNA-TAG labeling in HeLa lysates

We report the application of RNA-sequencing to streptavidin-affinity purified RNA transcripts from HeLa cell lysates. RNA was biotinylated using the enzymatic RNA-TAG methodology using two variants of E. Coli TGT, the wild type enzyme and an obligate dimeric form of the enzyme for comparison of selectivity. From this analysis, it was determined that the obligate dimer enabled affinity purification of the desired transcript with much greater selectivity. Overall design: Examination of 2 different RNA labeling conditions in HeLa cell lysates