GSM7653405: U2OS_CHR, BMAL1, 16h, ChIP; Homo sapiens; ChIP-Seq
Pipeline of  ChIP  Enrichment (1) DNA-protein cross-linking by formaldehyde. (2) Add Lysis Buffer to get whole cell lysate. (3) Lysate sonication (100-500bp). (4) Immunoprecipitation: Add primary antibody and beads to samples. (5) Elution and reverse cross-links. (6) qPCR for Quality Control. (7) Ready for  ChIP -seq library construction. 1 Sample QC The corresponding sample quality control protocol will be selected depending upon sample types and product requirements. 2 End Repair, add “A”, Adaptor Ligation After the reaction system and program are configured and set up, DNA fragments obtained from ChIP are subjected to end-repair, and then a single 'A' nucleotide is added to the 3' ends of the blunt fragments. The reaction system and program for adaptor ligation are subsequently configured and set up to ligate adaptors with the DNAs. 3 PCR The PCR reaction system and program are configured and set up to amplify the product. 4 Library QC The corresponding library quality control protocol will be selected depending upon product requirements. 5 Circularization Single-stranded PCR products are produced via denaturation. The reaction system and program for circularization are subsequently configured and set up. Single-stranded cyclized products are produced, while uncyclized linear DNA molecules are digested. 6 Sequencing Single-stranded circle DNA molecules are replicated via rolling cycle amplification, and a DNA nanoball (DNB) which contain multiple copies of DNA is generated. Sufficient quality DNBs are then loaded into patterned nanoarrays using high-intensity DNA nanochip technique and sequenced through combinatorial Probe-Anchor Synthesis (cPAS).