GSM6030637: MPRA-Plasmid-Library; Homo sapiens; OTHER
250ng of plasmid library DNA was used for the construction of sequencing libraries Library construction was performed using a two-step PCR amplicon approach. First amplification was performed at 98°C for 30” followed by ten cycles of 98°C for 10”, 71°C for 30”, 72°C for 30”, and final elongation at 72°C for 5' with Forward primer: 5'-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGgccagcgccaggatcaac -3' and Reverse Primer: 5'- GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGcgccccgactctagctagac -3' using Q5 Polymerase. PCR product was purified using 1.8X Ampure XP beads. Second amplification was was performed at 98°C for 30” followed by eight cycles of 98°C for 10”, 67°C for 30”, 72°C for 30”, and final elongation at 72°C for 5' with Forward primer: 5'- aatgatacggcgaccaccgagatctacactcgtcggcagcgtc -3' and Reverse Primer: 5'- caagcagaagacggcatacgacattcgccttagtctcgtgggctcgg -3' using Q5 Polymerase. PCR product was purified using 1.8X Ampure XP beads.