GSM4980050: pasRNA_ABAT_replicateB (DMSO); synthetic construct; OTHER
The ABAT PAS RNA used in SHAPE-MaP experiment was synthesized by MAXIscript™ SP6/T7 Transcription Kit according to manufacturer's protocol. After purification, 500 ng ABAT PAS RNA for each SHAPE sample was refolded before being treated with DMSO or 100 mM 1-Methyl-7-nitroisatoic anhydride (1M7) for 75 second. RNA was then isolated and subject to SHAPE-MaP reverse transcription with gene specific primer. Second-strand synthesis was performed with Q5 hot start high-fidelity DNA polymerase. Deep sequencing was performed by Illumina MiSeq PE250.