GSM5289538: C2C12-Mm-MUNC-genomic-IC-5NIA; Mus musculus; OTHER
Cells were grown to ~80% confluency and washed briefly twice with ice-cold PBS. Total RNA for in-cell treatments was extracted by Trizol reagent (Invitrogen #15596018) and purified with column-based Direct-zol RNA Miniprep kit (Zymo Research #R2051), while cell-free samples were prepared according to SHAPE-MaP protocol. MUNC amplicons were generated by gene-specific MaP reverse transcription and PCR. cDNA products were converted to libraries using Illumina Nextera XT library prep kit and sequenced on an Illumina MiSeq instrument.