| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
| zygote Hi-C R2 |
In Situ Hi-C library generation was performed with some modification accordingto the procedures described in detail elsewhere (Rao et al., 2014). All samples were crosslinked with 1%formaldehyde at room temperature for 10 minutes. Nuclei were permeabilized. DNAwas digested with 10 units of MboI, and the ends of restriction fragments werelabeled using biotinylated nucleotides and ligated in a very small volume. Thecrosslinks, ligated DNA was pelleted, washed with 10mM Tris buffer once, thenreversed in 10mM Tris buffer and 1.25mg/ml proteinase K (Qiagen, 19133),incubated at 65°C for 5h and 75°C for 30min to inactivate the protease. The reversedDNA was sheared to a length of ~400bp, and directly treated with EndRepair/dA-Tailing Module (NEB, E7442L) and Ligation Module (NEB, E7445L).The biotin-labelled ligation junction DNA was pulled down with streptavidin beadsand amplified for 12-14 cycles. The amplification mixture was purified usingAMPURE XP beads to select size 400-600bp. The result |
OTHER |
GENOMIC |
unspecified |
PAIRED
|
|
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