| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
|
Cells were lysed with 6 μl lysis buffer for 10 min on ice. Then, we added 5 μl ddH2O, 4 μl 5 xTTBL, 5 μl TTE mix V5 (TD502, Vazyme) to the tube and the mixture were incubated at 37C for 30 min. The 5 μl of 5 xTS buffer (TD502, Vazyme) was next added and incubate at the room temperature for 5min to terminate the reaction. Next, we added 40 ng carrier RNA (59824, QIAGEN), 73 μl TE (Tris EDTA), 100 μl phenol chloroform to the reaction product. After vortexing and incubating for 3 min at the room temperature, the product was transferred to a phase lock tube (WM5 2302820, TIANGEN) and centrifuge at 12,000 rpm for 5 min. The supernatant was then transferred to a new 1.5ml tube and added 650 μl ethanol, 24 μl NaOAc (3M), 2 μl glycogen for DNA precipitation at 20 °C overnight. Then, DNA pellet was resuspended in 29μl ddH2O and added with 10 μl 5 xTAB, 1 μl TAE (TD502, Vazyme), 5 μl N5XX primer, 5 μl N7XX primer (TD202, Vazyme) for PCR reaction. DNA was amplified using the following cycling protocol: 72 °C for 3min; 98 °C for 30s; 12 cycles of 98 °C for 15s, 60 °C for 30s and 72 °C for 3min; 72 °C for 5min. After amplifying, libraries were performed size selection with 0.5-1.5 x AMPure beads (A63882, Beckman) and were sequenced with 150 bp paired end reads on an Illumina NovaSeq 6000 platform. |
ATAC-seq |
GENOMIC |
RANDOM |
PAIRED
|
|
