GSM8751682: HS/AAV6 + ANAK - Sample6; Homo sapiens; OTHER
Genomic DNA was isolated with QIAamp DNA Micro Kit (QIAGEN) PCR amplicons for individual samples were generated by nested PCR starting from >50–100 ng of purified gDNA. The first PCR step was performed with GoTaq G2 DNA Polymerase 1007 (Promega) according to the manufacturer's instructions using the following amplification protocol: 95°C for 5 min, (95 °C for 0.5 min, 60 °C for 0.5 min, 72 °C for 0.25 min) × 20 cycles, 72 °C for 5 min. The second PCR step was performed with GoTaq G2 DNA Polymerase (Promega) according to manufacturer's instructions using 5 μl of the first-step PCR product and the following amplification protocol: 95 °C for 5min; 95°C x 0.5 min, 60°C x 0.5 min, 72°C x 0.3 min, (20cycles); 72°C x 5 min. Second-step PCR primers were endowed with tails containing P5/P7 sequences, i5/i7 Illumina tags to allow multiplexed sequencing and R1/R2 primer binding sites. The PCR amplicon from each sample was separately purified by using a QIAquick PCR Purification kit (Qiagen).