We were very happy with the method. It gave good results in the end, and required much smaller samples than we need to reliably perform conventional ChIP-seq.
In our view, the main advantages of the ChIPmentation kit compared to our conventional ChIP-seq protocol are (most important first):
- smaller sample requirement,
- simpler workflow with less that can go wrong,
- slightly higher resolution and signal: noise ratio.
ChIPmentation sequencing profiles for p65. Chromatin preparation and immunoprecipitation have been performed on stimulated NIH3T3 cells using the iDeal ChIP-seq kit for TFs (Cat. No. C01010055). Chromatin from 4,000,000 cells was used for the immunoprecipitation in combination with either anti-p65 antibody or IgG. The library preparation was performed with the TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031).
TAG Kit for ChIPmentation
Tagmentation is a reaction where an enzyme (a transposase) cleaves DNA and incorporates sequencing adaptors at the ends of the DNA fragments in one step. In our ChIPmentation technology we combine chromatin immunoprecipitation and tagmentation in one streamlined workflow where the tagmentation step occurs directly on chromatin. The TAG Kit for ChIPmentation has been developed for researchers who would like to perform ChIPmentation using their own validated ChIP protocol. Our TAG Kit for ChIPmentation includes reagents for tagmentation-based library preparation and is compatible with any ChIP protocol based on magnetic beads. The primer indexes for multiplexing must be purchased separately and are available as a reference: 24 SI for ChIPmentation, Cat. No. C01011031.
TESTIMONIALOne of our issues was that we could obtain only a limited number of cells, which is not enough for canonical ChIP-seq protocols. To solve this issue, we used the Diagenode ChIPmentation solution composed of iDeal ChIP-seq Kit for Transcription Factor, TAG Kit for ChIPmentation, and 24 SI for ChIPmentation. We performed ChIPmentation with IP-Star automated system for GATA6 in 2 million GATA6-overxpressing iPS cells. The result showed clear signal/noise ratio and was highly reproducible. This solution also worked in vitro differentiated definitive endoderm cells (data not shown).
Region 1
Region 2
Figure 1. ChIPmentation sequencing profiles for Gata6
Chromatin preparation and immunoprecipitation have been performed on hiPSCs (human induced Pluripotent Stem Cells) overexpressing Gata6 using the iDeal ChIP-seq kit for TFs (Cat. No. C01010055). Chromatin from 2,000,000 cells was used for the immunoprecipitation in combination with either anti-GATA6 antibody. The library preparation was performed with the TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031).
- Characteristics
Comparing to classical ChIP-seq, ChIPmentation enables:
- An easier protocol
- Faster time to results
- Flexibility – the TAG Kit for ChIPmentation combained with 24 SI for ChIPmentation allows you to perform ChIPmentation with any ChIP protocol (based on magnetic beads)
ChIPmentation on histone marks using TAG KIT for ChIPmentation
ChIPmentation sequencing results for four histone marks. Chromatin preparation has been performed on 7M K562 cells using the Auto iDeal ChIP-seq Kit for Histones (Cat. No. C01010057), TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031)*. Diluted chromatin from 1,000,000 cells was used for the immunoprecipitation with the Diagenode antibodies targeting H3K4me1 (Cat. No. C15410194), H3K27me3 (Cat. No. C15410195), H3K27ac (Cat. No. C15410196) and IgG (Cat. No. C15410206). The experiment was performed on the IP-Star® Compact Automated System.
*The Auto iDeal ChIP-seq for Histones (C01010057), TAG Kit for ChIPmentation (C01011030) and 24 SI for ChIPmentation (C01011031) form together the Auto ChIPmentation Kit for Histones* (C01011000).
ChIPmentation on non-histone proteins using TAG Kit for ChIPmentation
ChIPmentation sequencing results for two non histone factors. Chromatin preparation has been performed on 25M K562 cells using the iDeal ChIP-seq kit for TFs (Cat. No. C01010055). Diluted chromatin from 1,000,000 or 4,000,000 cells was used for the immunoprecipitation with the Diagenode antibodies targeting CTCF (Cat. No. C15410210) and RNA PolII (Cat. No. C15100055 ) respectively. The library preparation was performed with the TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031).
- 出版物
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A dataset of definitive endoderm and hepatocyte differentiations fromhuman induced pluripotent stem cells.
Tanaka Y. et al.
Hepatocytes are a major parenchymal cell type in the liver and play an essential role in liver function. Hepatocyte-like cells can be differentiated in vitro from induced pluripotent stem cells (iPSCs) via definitive endoderm (DE)-like cells and hepatoblast-like cells. Here, we explored the in vitro differentiation ...GATA6 is predicted to regulate DNA methylation in an in vitro model ofhuman hepatocyte differentiation.
Suzuki T. et al.
Hepatocytes are the dominant cell type in the human liver, with functions in metabolism, detoxification, and producing secreted proteins. Although gene regulation and master transcription factors involved in the hepatocyte differentiation have been extensively investigated, little is known about how the epigenome is... - 相关产品
