GSM6588975: Z4KDrep1; Drosophila melanogaster; ATAC-seq
After respective treatments, cells were harvested and processed according to experiment. Hi-C and ChIP-seq cells were crosslinked with 1% formaldehyde for 10 min followed by 5 min glycine treatment to quench crosslinking reaction, flash freezed and stored at -80°C. Cell pellets were resuspended in 50 μl Tn5 transposase mixture (2.5 μL Tn5, 25 μL 2X TD buffer, 22.5 μL RNase Free Water), and incubated at 37°C for 30 min. After the reaction, cells were cooled on ice and DNA was purified using the Minelute Kit (QIAGEN). Transposed DNA was eluted in 10 μl of Elution Buffer (10mM Tris, pH8.0) and used for real-time PCR amplification in the reaction mixture (2x KAPA HiFi mix and 1.25 μM indexed primers) using the following conditions: (1) 72°C for 5 min; (2) 98°C for 30 s; (3) 98°C for 10 s, (4) 63°C for 30 s, (5) 72°C for 1 min, (6) Repeat steps 3-4 for 4 times, (7) Hold at 4°C.