GSM3223916: GM12878 cell number 100; Homo sapiens; OTHER
Single cells were captured using the C1 Single-Cell Auto Prep IFC microfluidic chips and assayed using the transcript-indexed single-cell ATAC-seq protocol. In a 96-well plate, 6.5 µL from a total of 15ul of harvested libraries were amplified in a 20 µL PCR for an additional 17 cycles (1uM each V1 primer, 1x NEBnext High-Fidelity PCR Master Mix) using the following PCR conditions: 98°C for 30 s, and cycling at 98°C for 10 s, 63°C for 30 s, and 72°C for 45 s. Then 2ul of the previous V1 PCR reaction were transferred to a nested V2 PCR reaction with V2 primers at the same concentration and using same cycling parameters as the V1 reaction, for 15 cycles. Next 2ul of the previous V2 PCR reaction were transferred to a cell barcoding V3 PCR reaction with V3 primers at the same concentration and using the same cycling parameters as the V2 reaction, for 15 cycles. Finally V3 reactions were pooled, purified, sized selected via PAGE, and quantified by qPCR prior to sequencing.
GEO Accession
GSM3223916