GSM8774696: MEF, Supernatant1, Rep2; Mus musculus; OTHER
5 µL of Proteinase K + 20 mM EDTA was added to each supernatant sample, and each pellet was resuspended in 500 µL of 1% SDS Lysis Buffer + 5 µL of Proteinase K + 20 mM EDTA.  Pellet samples were then broken up with a 20-gauge syringe.  All supernatant and pellet samples incubated for 1 hour at 50°C.  1% SDS was added to each sample, and the pellet samples were again broken up with a 20-gauge syringe.  All samples were incubated overnight at 65°C.  DNA was purified via chloroform extraction and ethanol precipitation. i5 and i7 sequencing adaptors were directly added to DNA isolated from all supernatant samples with an NEBNext Multiplex Oligos for Illumina kit. DNA isolated from all pellet samples was digested with 1 uL of Tn5 protein at 37°C for 30 minutes at 1000 RPM, and i5 and i7 sequencing adaptors were added via PCR.