GSM3142637: 6_G1_HeLa; Homo sapiens; OTHER
Cells were resuspended in 10 mM Tris-HCl pH 8.0, 0.5% w/v SDS, 100 mM EDTA supplemented with 20 µg/ml RNase A and 0.2 mg/ml proteinase K and incubated at 50˚C for 1 h. The lysate was allowed to cool to room temperature and sample volumes were adjusted to 0.5 mL by addition of TE buffer. The samples were mixed with an equal volume of phenol:chloroform:isoamyl alcohol pH 8.0 (25:24:1) followed by single or double extraction using an equal volume of chloroform. DNA was then precipitated using isopropanol in the presence of potassium acetate. 500 ng of fragmented genomic DNA was used for library construction for Illumina sequencing. Indexed genomic DNA libraries were prepared using a NEBNext Ultra II library prep kit for Illumina without size selection, and NEBNext Multiplex Oligos for Illumina using four cycles of amplification, followed by two rounds of clean-up using Agencourt AMPure XP beads. Library samples were quantified by qPCR using a NEBNext Library Quant Kit for Illumina and a Rotor-Gene real time PCR cycler.
GEO Accession
GSM3142637