GSM5961955: HL-60_4C_DMSO, rep1; Homo sapiens; OTHER
Nuclei were purified from lysates of fixed HL-60 (7 milions cells). Chromatin was then digested with NlaIII enzyme and ligated. After DNA purification, the chimeric DNAs were digested with DpnII enzyme and re-ligated. DNA was purified to proceed the library preparation. 4C libraries were amplified using specific primers composed of P5/P7 Illumina sequence supplemented with indexes and sequences corresponding to the promoter of NFkB2 (viewpoint). The “Expend Long Template PCR System” kit (Roche) was used using 300 ng of the 4C library following the manufacturer's instruction. The following amplification parameters were used: denaturation for 2 min at 94°C followed by 30 cycles (94°C - 15 seconds, 58°C - 1 minute and 68°C – 3 minutes) and 7 min at 68°C. Libraries were purified and concentrated with ampureXP beads.