Chromatin Immunoprecipitation Sequencing by Ion Torrent Next-Generation Sequencing
Chromatin Immunoprecipitation Sequencing (ChIP-Seq)
Assessing protein-DNA interactions at genome-wide level is essential for understanding a large number of biological processes during normal development and disease. Isolation of protein-DNA interactions by chromatin immunoprecipitation (ChIP) followed by massively parallel DNA sequencing is an unbiased method for identifying and quantitating DNA sites that are associated to a protein(s) of interest. Of particular interest are DNA regions involved in gene regulation, nucleosome positions, and epigenetic mechanisms such as chromatin formation associated with histone modifications. Identifying transcription factor binding sites is important for understanding cell differentiation, environmental and drug responses, as well as alterations to these responses in disease states. Understanding the epigenetic modifications to chromatin is critical for determining the influence of chromatin states on transcription and the dynamic interplay between the epigenome and gene regulatory networks.
ChIP-Seq Uses Proven Methodology
ChIP-Seq is an in vivo assay that uses antibodies to capture proteins that are bound to DNA directly or that interact within DNA-bound protein complexes. The ChIP-Seq workflow begins with formaldehyde treatment of live cells to crosslink proteins to DNA; this is followed by cell lysis and fractionation of the chromatin. An antibody specific to the protein of interest is used to immunoprecipitate the crosslinked protein-DNA complexes. The crosslinks are then reversed, and the resulting purified DNA is used in subsequent DNA fragment library construction for massively parallel DNA sequencing. Following sequencing, the sequence reads are mapped to a reference sequence and counted to identify specific DNA regions bound by proteins.
Solutions for ChIP-Seq Analysis
ChIP-Seq analysis provides fundamental insights, at a genome-wide level, into how chromatin is organized and regulated and how gene transcription is altered during many biological processes and disease. Consider the following solutions to begin ChIP-Seq analysis in your lab:
- The Ion Proton™ System enables 2 to 4 ChIP-Seq reactions per run with the Ion PI™ Chip, and higher multiplexing with the Ion PII™ Chip*
- The MAGnify™ Chromatin Immunoprecipitation System enables a simplified construction of ChIP-Seq libraries in approximately 5 hours
- Dynabeads® are designed to reduce nonspecific binding and assay background and offer improved reproducibility via optimized magnetic DNA purification
- Reliable ChIP-Seq results depend on high-quality, specific antibodies; Life Technologies offers a number of antibodies qualified for ChIP
The Ion Proton™ System combined with the MAGnify™ Chromatin Immunoprecipitation System delivers fast, reliable, and high-quality ChIP-Seq results at an affordable price.
*The content herein may relate to products that have not been officially released and is subject to change without notice.
For Research Use Only. Not for use in diagnostic procedures.