Overview

Chromatin immunoprecipitation sequencing (ChIP-seq) is a method used to identify and map global interactions of the genome with DNA binding-proteins involved in gene regulation or chromatin organization. In a typical ChIP workflow, genomic DNA that has been temporarily bound to proteins by crosslinking is sheared and the bound DNA fragments are selectively enriched by immunoprecipitation. Once purified, the DNA fragments are used in the construction of a library for sequencing. The resulting sequences will provide information on binding sites for the proteins of interest.

Workflow

The amount of enriched DNA obtained following purification is generally very low. ThruPLEX DNA-seq Technology is being used by many leading laboratories to construct libraries used in CHIP-seq because of its superior performance with low input amounts of DNA.

ThruPLEX Technology*

ThruPLEX TechnologyThruPLEX is a three step process that is performed in a single tube.

Step 1.ThruPLEX technology features improved repair chemistry.

Step 2: The background is reduced using double-stranded adaptors with no single-stranded tails. Blunt end ligation occurs with high-efficiency blunt-end ligation Blocked 5’ends reduce adaptor-adaptor ligation.

Step 3. Background is reduced by destroying unused adaptors after ligation.

Overall, the time to complete the process is reduced by using compatible buffers and multiplexed enzymatic reactions that do not require intermediate purifications.

Request our application note: Small cell number ChIP-seq using ThruPLEX® DNA-seq Kit as a tool for epigenetic profiling

Learn more about ThruPLEX DNA-seq Kit.
*Protected by US patent 7,803,550 and EP1924704

Workflow

 

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A basic workflow for Chip-seq includes a ThruPLEX library preparations step.