SHAPE-MaP of Ty3 gRNA: in vivo treated with 50 mM vanillin NAI amplicon 1 rep2
Yeast cells in mid-log phase were treated with 50 mM vanillin for 30 min. After this step cells were subjected to SHAPE modification procedure (100 mM NAI in DMSO/20 min). SHAPE-modified RNA was extracted, 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.