title: GSM8510969: HS187,HS188,HS189, Human sperm, scHi-C; Homo sapiens; Hi-C
library_name: GSM8510969
library_construction_protocol: All animal experimental procedures were carried out following the Institutional Animal Care and Use Committee (IACUC) guidelines of Peking University. 10–12-week-old C57BL/6J male mice were purchased from Peking University Laboratory Animal Centre. The single-cell preamplification product was diluted with water to 5 ng/uL. 2 μL of each diluted sample was transposed by the addition of 3 μL Transposition Mixture (1 μL 5 X TTBL, 0.25 μL TTE Mix V50 (Vazyme TD501), 1.75 μL water) and incubated at 55 °C for 10 min. Transposition was stopped by adding 1.25 μL 0.2% SDS and then the plate was incubated at room temperature for 10 min. Then, 2 μL 5 μM i5 unique dual (UD) index and 2 μL 5 μM i7 unique dual (UD) index were added in each well. Next, the sperm Hi-C library was amplified for 10 cycles of PCR with 9.75 uL KAPA PCR mix (Roche, KK2102) (4 uL 5× KAPA HiFi GC Buffer, 0.6 uL dNTP Mix (10 mM each), 4 uL KAPA HiFi DNA Polymerase (1 U/uL) and 4.75 uL water). The barcoded single-cell libraries were then pooled and purified with 0.6× and 0.15× VAHTS DNA Clean Beads (Vazyme, N411-02). The final libraries were sequenced with paired-end 150bp reads on a NovaSeq X plus (Illumina) platform according to the manufacturer's instructions.