title: GSM4237865: 55_Wild-type_integrase_HepG2_DNA_rep2; Homo sapiens; OTHER
library_construction_protocol: The cells were washed with PBS three times, and genomic DNA and total RNA was extracted using AllPrep DNA/RNA mini kit (Qiagen). Messenger RNA (mRNA) was purified from the total RNA using Oligotex mRNA mini kit (Qiagen) and treated with Turbo DNase to remove contaminating DNA. For each RNA replicate, 3x500ng was reverse transcribed with Superscript IV (Invitrogen) using a primer downstream of the barcode, which contained a UMI. The resulting cDNA was pooled and split into 8 reactions, amplified with Kapa Robust polymerase for 3 cycles using an indexed P5 primer and P7. PCR products are then cleaned up with AMPure XP beads (Beckman Coulter) to remove the primers and concentrate the products. These products underwent a second round of amplification in 6 reactions per replicate and followed in real time with Sybr Green. All reactions were pooled at this point, cleaned up with AMPure CP beads, run on an agarose gel, and submitted for sequencing.
attributes.tag: GEO Accession
attributes.value: GSM4237865