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Hi-C library was prepared according to the protocol of the commercial Hi-C library construction kit (ProxiMetaTM KIT, USA). Briefly, for each sample, 100mg was suspended in 1mL of crosslinking solution and incubated for 15 min at room temperature with occasional mixing gently and the reaction was stopped by adding 100 microliter quenching Solution for 20 min at room temperature incubation. Cells in the sample were suspended and lysed by lysis buffer, and purified by Recovery Beads. The pure sample DNA was then fragmented by Fragmentation Enzyme with 1 hr at 37 degrees Celsius. Digest DNA was then ligated by ligation enzyme at 16 degrees Celsius overnight and then the ligated DNA was purified by magnetic beads. After purification, biotinylation was done by streptavidin beads and then processed into the sequencing library according to the protocol. The concentration of DNA was measured by Qubit dsDNA HS Assay Kit and the distribution of the library was assessed by bioanalyzer or similar instrument. Hi-C libraries were then prepared and sequenced (2*150 bp, Illumina NextSeq, 10 first bases as index). |
Hi-C |
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PCR |
PAIRED
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