GSM7846629: iPSC_DNA_rep2; Homo sapiens; OTHER
Genomic DNA and total RNA were purified from the infected cells using an AllPrep DNA/RNA mini kit (Qiagen). To remove contaminating DNA, RNA was treated with Turbo Dnase (Thermo Fisher Scientific). 120 ug total RNA samples were reverse-transcribed with SuperScript II (Invitrogen) using a barcode-specific primer, which has a 16-bp unique molecular identifier (UMI). The cDNA and 48 μg genomic DNA from each sample were used for 3-cycle PCR with specific primers that add sample index and UMI. A second round PCR (21 and 26 cycles for DNA and RNA barcode samples, respectively) was performed using P5 and P7 primers. PCR products were size-selected with an agarose gel, purified with AmPure XP beads (Beckman coulter) and pooled. Custom DNA sequencing; 15-cycle paired-end reads, 16-cycle index read1 for UMI, and 10-cycle index read2 for sample index.