GSM4504483: SHAPE-Map data denaturing control in in-cell condition; Homo sapiens; OTHER
From each sample of RNA from Hela in-cell and cell-free probing experiments, 1-3 µg were subjected to mutational profiling (MaP) reverse transcription using a primer specific to CCTT. The cDNA generated was buffer exchanged over Illustra microspin G-50 columns (GE Healthcare). Output cDNA (5 µL) was used as a template for 50 µL PCR reactions (Q5 Hot-start polymerase, NEB) with primers made to amplify CCTT and add adapter sequences [1x Q5 reaction buffer, 250 nM each primer, 200 µM dNTPs, 3% DMSO, 0.02 Units/µL Q5 Hot-start polymerase]. PCR proceeded in a touchdown format and high-throughput sequencing library construction.
GEO Accession
GSM4504483