title: GSM8644386: Plk1as_3MB_rep1; Homo sapiens; ATAC-seq
library_construction_protocol: 50,000 cells were resuspended in 50 uL transposition reaction mix containing 25 uL Tagment DNA buffer, 16.5 uL 1x PBS, 0.5 uL 10% Tween-20, 0.5 uL 1% Digitonin, 2.5 uL Tn5 Transposase, and 5 uL nuclease-free water. Reaction mixture was incubated at 37°C for 30 min in a Thermomixer set at 1000 rpm. DNA was isolated using a QIAquick PCR Purification kit according to manufacturer's protocol, and DNA was eluted in 10uL of the supplied elution buffer. The entirety of the purified DNA was added to a PCR reaction containing 10 uL nuclease-free water, 2.5 uL each of 25uM NextEra i5 and i7 barcoded primers, and 25 uL NEBNext high-Fidelity 2X PCR Master Mix. Samples were amplified on the following program: 72°C 5 min, 90°C 30 sec, (98°C 10 sec, 63°C 30 sec, 63°C 1 min) with certain number of cycles. 5 uL of the PCR reaction was used to calculate the number of cycles needed for the libraries. Amplified samples were size selected using SPRIselect beads by doing a 0.5x standard bead cleanup followed by a 1.8x standard bead cleanup. Samples were eluted from the beads using 20uL of nuclease-free water.
identifiers.namespace: GEO
identifiers.id: GSM8644386_r1