title: GSM3143116: scMNase-seq_NIH3T3_MNase0.1U_GB4007080; Mus musculus; MNase-Seq
library_construction_protocol: 0.1 units of MNase was added to 5 million of NIH/3T3 cells in 1ml of cell lysis buffer (10mM Tris-HCl, pH 7.5, 10mM NaCl, 3mM CaCl2, 0.1% Triton X-100) and incubated at 37°C for 5 min. The reaction was stopped by adding 5mM of EDTA and spin down. Wash once with FACS buffer. Single cell were sorted by flow cytometer into 100ul of Stop Buffer (10mM Tris-HCl, pH 7.5, 10mM NaCl, 0.15% SDS, 10mM EGTA) includ 1 µl of 20mg/ml Proteinase K. Following incubation at 65°C for overnight, 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.
attributes.tag: GEO Accession
attributes.value: GSM3143116