title: GSM2906296: scMNase-seq_mESC_GB4394; Mus musculus; MNase-Seq
library_construction_protocol: For Micrococcal Nuclease (MNase) digestion, 8 ml diluted MNase (1:3000 in lysis buffer) was added to the sorted single cells and incubated at 37°C for 5 min. The reaction was stopped by addition of 80 ml of Stop Buffer (10mM Tris-HCl, pH 7.5, 10mM NaCl, 0.15% SDS, 10mM EGTA), 1 ml of 20mg/ml Proteinase K and 5 ml of 6ng/ml circular carrier DNA. Following incubation at 55°C for 1 hour, DNA was purified by Phenol-chloroform extraction and precipitation with ethanol in the presence of glycogen. Universal adaptors were then ligated to the DNA ends as described by Jin et al (Nature 2015) . The libraries were amplified using indexed primers with the PCR condition: 98°C, 10”; 67°C, 30”; 72°C, 30” for 23 cycles. The fragments between 200bp to 300bp were isolated on E-gel and subjected to paired-end sequencing on Illumina HiSeq2500.
identifiers.namespace: GEO
identifiers.id: GSM2906296