ENA first public
2022-02-22
ENA last update
2022-02-22
External Id
SAMEA8920608
INSDC center alias
Tianjin Medical University
INSDC center name
Tianjin Medical University
INSDC first public
2022-02-22T00:26:47Z
INSDC last update
2022-02-22T00:26:47Z
INSDC status
public
Submitter Id
MT E6.5 Epi ATAC-seq
common name
house mouse
geographic location (country and/or sea)
China
isolate
E6.5 Epi
protocol
10 E6.5 Epi 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 μl ddH2O, 10 μl 5×TTBL, 3 μl TTE mixV50 (TD501, Vazyme)) and gently flicked. The reaction was incubated at 37°C for 0.5 h in a PCR cycler, before 1 μl 0.5M EDTA, 1 μl proteinase K (20 mg/ml) and 5 μl 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 μl ddH2O. 15 μL 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 μL) Ampure XP beads were added to remove fragments above 1 Kb and another 0.5 volume (25 μL) Ampure XP beads were added to collect the 200-1,000bp fragments for sequencing.
receipt date
2021-06-18
sample name
MT E6.5 Epi ATAC-seq