Low cell ChIP-Seq library preparation kit

The low cell ChIP-Seq kit from Active Motif provides a complete ChIP-Seq workflow for chromatin preparation, immunoprecipitation and NGS library preparation from limited amounts of cells or small tissue biopsies.

This assay is robust enough to work with both high abundance histone modifications and low abundance transcription factor proteins. The Next Gen DNA Library and Next Gen DNA Indexing reagents are included to generate high complexity sequencing libraries that are compatible with Illumina® platforms.

Benefits of the Low Cell ChIP-Seq kit
• As little as 1,000 to 50,000 cells needed depending on protein abundance
• Complete workflow includes chromatin preparation, immunoprecipitation and Illumina-compatible NGS library prep
• Improved signal-to-noise ratio for better peak calling and lower background
• Molecular identifiers (MIDs) incorporated within the P5 adapter

The Low Cell Chip-Seq kit produces high quality ChIP-Seq peaks

The Low Cell ChIP-Seq Kit was used to generate ChIP-Seq data from 10,000 MDA-MB-468 cells using an antibody directed against bromodomain-containing 4 (BRD4) protein, or 5,000 GM12878 cells using antibodies directed against histone H3K27ac or histone H3K27me3. Following ChIP, Illumina-compatible sequencing libraries were prepared using the included Next Gen DNA Library Kit and Next Gen Indexing Kits and sequenced using the NextSeq 500. Results show high quality ChIP-Seq peaks can be obtained using as few as 5,000 cells for both active and repressive histone modifications and as few as 10,000 cells for BRD4. Peaks were compared to ENCODE data sets using 20 million cells.

The Low Cell ChIP-Seq kit produces nice ChIP peaks for both robust and low abundance target proteins

Summary of Low Cell ChIP-Seq Kit data sets using both robust and low abundance target proteins. Nice peaks above background were observed from as few as 1,000 cells for the histone modifications H3K4me3, H3K4me1, and H3K27me3. Both active and repressive histone modifications are shown, indicating that the method is sensitive enough to obtain good ChIP-Seq signal even when working with repressive chromatin. Lower abundance proteins, including BRD4 and insulator protein CTCF showed nice ChIP enrichment from as few as 10,000 cells. Results shown are from a single Low Cell ChIP-Seq Kit reaction, however, data has been reproducibly generated across multiple experiments.

Material available for download
Low cell ChIP-Seq kit manual
Chromatin sonication recommendations for ChIP

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