Determining how proteins interact with DNA to regulate gene expression is essential for understanding many biological processes and disease states. DNA-protein interaction sites can be isolated by chromatin immunoprecipitation (ChIP). Illumina combines whole-genome ChIP with massively parallel DNA sequencing to identify and quantify binding sites for DNA-associated proteins.
Illumina's TruSeq ChIP sample prep kit protocol enables researchers to cost-effectively and reliably identify binding sites for a broad range of targets across the entire genome with high resolution and without constraints. The streamlined workflow reduces the number of purification, sample transfer, pipetting and clean-up steps. TruSeq universal adapters enable robust multiplex sequencing.
The TruSeq ChIP sample prep process begins with the enrichment of specific cross-linked DNA-protein complexes using an antibody against a protein of interest (A-B). The stretches of DNA bound to the target protein are then isolated and used as input DNA for library generation. DNA fragments are end-repaired and an 'A'-base added to the blunt ends of each strand, preparing them for ligation to the sequencing adapters (C-D).
Each TruSeq adapter contains a 'T'-base overhang on the 3'-end providing a complementary overhang for ligating the adapter to the A-tailed fragmented DNA (E). Final product is created (F) and after size selection, all of the ChIP DNA fragments can be simultaneously sequenced (single-read or paired-end) on Illumina sequencers.
TruSeq ChIP Sample Prep Kit Highlights