ENA first public
2022-02-22
ENA last update
2022-02-22
External Id
SAMEA8550505
INSDC center alias
Tianjin Medical University
INSDC center name
Tianjin Medical University
INSDC first public
2022-02-22T00:26:34Z
INSDC last update
2022-02-22T00:26:34Z
INSDC status
public
Submitter Id
WT-2i ATAC-seq rep1
common name
house mouse
geographic location (country and/or sea)
China
geographic location (region and locality)
TianJin
isolate
mESCs
protocol
50,000 mES cells were used for ATAC-seq. After washed by PBS, the cells or epiblasts were resuspended in lysis buffer (10 mM Tris-HCl, pH 7.5, 10 mM NaCl, 3 mM MgCl2, 0.1%(v/v) igepal CA-630 and 1x PIC). After 10 min incubation on ice, cells were spinned down by centrifugation at 500g, 4～C. Then the pellets were resuspended in transposome mix (37 ml ddH2O, 10 ml 5℅TTBL, 3 ml TTE mixV50 (TD501, Vazyme)) and gently flicked. The reaction was incubated at 37～C for 0.5 h in a PCR cycler, before 1 ml 0.5M EDTA, 1 ml proteinase K (20 mg/ml) and 5 ml 10% SDS were added to stop the reaction. After another incubation at 50～C for 1 h, DNA was purified with PCI and resuspended with 30 ml ddH2O. 15 mL DNA were amplified with i5 and i7 primers (Vazyme, TD202) and KAPA 2℅PCR mix (KM2602) following the cycling program with heated lid, 72～C, 5min; 98～C, 30s; 10 cycles for 98～C, 10s; 60～C, 30s; 72～C,30s; and 72～C for 5min, hold at 4～C. Upon the program completed, 0.5 volume (25 mL) Ampure XP beads were added to remove fragments above 1 Kb and another 0.5 volume (25 mL) Ampure XP beads were added to collect the 200-1,000bp fragments for sequencing.
receipt date
2021-04-11
sample name
WT-2i ATAC-seq rep1