SHAPE-MaP of Ty3 gRNA: ex vivo NAI amplicon 1 rep1
Yeast cells in mid-log phase were harvested and RNA was extracted, denatured and refolded. After this step RNA was subjected to SHAPE modification procedure (100 mM NAI in DMSO/15 min). SHAPE-modified RNA was reverse transcribed with specific primer in Mn2+ conditions, and dsDNA was made by amplicon PCR. Final amplicon library was prepared by fragmentation and adapter ligation.