GSM6387253: Young, HSCs, CTCF, CUT&RUN; Mus musculus; OTHER
Fus low and Fus high HSCs(lineage-, c-kit+ Sca-1+ CD34-) from bone marrow of mice using fus gtp; Young(2 months) and Aged(30 months) HSCs(lineage-, c-kit+ Sca-1+ CD34-) from bone marrow of mice CUT&RUN: Young HSCs and Aged HSCs were sorted from 2-month mice and 30-month mice. 100,000 HSCs were centrifuged at 1,000g for 5min at 4℃, and washed with 200uL PBS.le. Then the conA beads were separated by magnet and resuspended with 200 μl Antibody Buffer (2mM EDTA, 0.02% digitonin and CTCF antibody (1:250, EMD Millipore, 07-729) in washing buffer)》 Next, DNA was digested using 200 μL pA-MNase Buffer. Next, Endprep reaction system(25uL DNA and H2O, 1.5 μLNEB Next Ultra II Endprep Enzyme Mix, 3.5uLNEB Next Ultra II Endprep Reaction Buffer) was subjected to PCR amplification. After amplification, the samples were purified with 0.3× -0.6× VATHS Beads and resolved with ddH2O to prepare for sequencing library after the DNA concentration was quantified with Qubit. The libraries were sequenced using an Illumina Hiseq platform with 150-bp paired-end.