GSM6895292: 4C_AHI1-ex20vp; Homo sapiens; OTHER
Circular chromosome conformation capture (4C) was done essentially as described (PMID: 22961246) using two million cells and two rounds of restriction digestion with either BglII and NlaIII, or with NlaIII and Csp6I, depending on the viewpoint, each time followed by ligation. First PCR 98°C for 30 sec, 10 cycles with 98°C for 15 sec, 61°C, 60°C or 54°C depending on the viewpoint for 20 sec with 0.5°C touch-down per cycle, 72°C for 2 min, then 30 or 25 cycles with 98°C for 15 sec, 56°C, 55°C or 49°C, depending on the viewpoint  for 20 sec, 72°C for 2 min, finally followed by 72°C for 1 min. Second PCR with 95°C, 3 min (initial melting) and 95°C, 20 sec, 62°C, 15 sec, 72°C, 40 sec (8 cycles). 4C sequencing libraries were generated in two PCR steps. In the first step, the  second ligation products, inverse primers and Q5 high fidelity enzyme were used. Libraries were generated in the second step with DNA from the first step essentially according to (PMID: 29224141) under real-time conditions.