In eukaryotes, chromatin is a complex composed of DNA and protein. It is of great significance to study the interaction between DNA and protein in chromatin form for understanding the gene expression mechanism of eukaryotes. As a classical method for protein-DNA interaction studies, Chromatin Immunoprecipitation (ChIP) is widely used in the researches of histone-specific modification sites and transcription factor binding sites. ChIP-Seq, which integrates chromatin immunoprecipitation and high-throughput sequencing technology, is another technological breakthrough in the study of protein-DNA interactions following ChIP-chip.
ChIP-Seq uses specific antibodies to immunoprecipitate the target protein, isolate the genomic DNA fragment bound to the target protein for purification and library construction, and then search for the DNA bingding site of the target protein in a genome-wide range through high-throughput sequencing, so as to obtain DNA fragment information that interacts with histones or transcription factors in a genome-wide manner.
Advantages
● Overall ChIP-Seq services from pre-ChIP experiments to high-throughput sequencing and bioinformatics analysis
● More than 20 histone modifications accessible
● Highly sensitive and highly specific antibodies for efficient enrichment of DNA fragments
Bioinformatics Analysis
RiboBio can provide multiple bioinformatics analysis services including bascis and advanced analysis services, welcome to contact order@ribobio.com to know more about it.
FAQs
1. What are the application scope of ChIP-Seq?
ChIP-Seq data is the result of DNA sequencing, providing researchers with the resources to further explore biological information in the following projects:
1) To determine what histone modifications will occur at a particular position in the DNA strand;
2) To detect the precise localization of binding sites of RNA polymerase II or other transcription factors on the genome;
3) To study the relationship between histone covalent modification and gene expression.
2. What are the sample requirements for ChIP-Seq?
ChIP-Seq needs to provide ≥10 ng of ChIP-enriched DNA samples with no protein, RNA or visible contamination. The DNA fragment size is in the range of 100-500bp, with a main peak of about 200bp being obvious. A test electropherogram after DNA interruption is needed to determine if the DNA fragment size meets the requirements, and a detailed sample information sheet and qPCR verification results after ChIP should also be attached.
3. Is PCR amplification required during sample preparation? Will the final result be affected by PCR amplification?
The amount of DNA samples obtained by ChIP is usually very limited, so PCR amplification is generally required in the library preparation process in order to obtain sufficient DNAs for experiment. If the amount of DNA sample supplied by the customer is sufficient, the repeatition of PCR can be reduced or cancelled. PCR amplification may lead to the deviation of sequencing and analysis results.
