GSM7070764: Pep13, HEK293T WT, GFP high; Homo sapiens; OTHER
Genomic DNA was isolated from the cell populations using QIAGEN QIAamp DNA mini kit or Machery Nagel Nucleospin Tissue kit. For Pep13, genomic DNA from unsorted input cells was also extracted. In addition, RNA was also isolated using Trizol for Pep13 cells. Genomic DNA was eluted in 50 uL of elution buffer. Libraries were then amplified using PCR with custom Illumina adapters, using Q5 high-fidelity PCR mix with 1000 ng input gDNA per sample. Libraries were amplified for a total of 24-27 cycles. After amplification, libraries were cleanup up using SPRISelect beads at a ratio of 0.7x. Purified library size was confirmed via gel and libraries were quantified using the KAPA qPCR Illumina library quantification kit. Libraries were subsequently pooled in a ratio based on the number of total cells collected from each sample. Pep13 RNA was reverse transcribed into cDNA and then preped using the same protocol described above.