ChIP-seq
ChIP-seq library construction typically involves the following steps: DNA fragmentation, end repair, single A-addition, adapter ligation, size selection, gel purification, and PCR amplification using platform-specific primers. During PCR amplification, it is important to avoid overamplification to prevent signal loss, bias, or redundancy. The quantity of PCR products should be sufficient for sequencing, with visible bands on an agarose gel with ethidium bromide staining. Libraries can be sequenced using single-end or paired-end strategies, with paired-end sequencing offering advantages such as increased coverage and improved alignment efficiency. The number of sequencing reads required depends on factors like antibody affinity and the number of target sites in the genome.