GSM8078779: Panc1 shRBM42-1 MYC 5UTR DMS bio rep 2; Homo sapiens; OTHER
RNA was purified using Trizol-chloroform as described above. To remove any contaminating genomic DNA, the RNA was treated with TurboDNAse per the manufacturer's instructions and repurified. Reverse transcription of the endogenous MYC 5'UTR was done with Induro Reverse Transcriptase (NEB) and a reverse primer targeting the coding sequence proximal to the ATG. The cDNA was purified was SPRI select beads (2x volume). The cDNA was amplified using 5'UTR Myc specific forward and reverse primers using PrimeStar Max (Takara) for 35 cycles using ~20% of purified cDNA product. The resulting DNA product was gel purified. Sequencing libraries were generated using the NEBNext Ultra II Kit following the manufacturer's instructions.