|TAG Kit for ChIPmentation MANUAL|
Diagenode’s TAG Kit for ChIPmentation in combination with the set of indexes (24 SI for ChI...
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.
One 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 2Takahiro Suzuki, Ph.D., Senior Research Scientist, Cellular Function Conversion Technology Team, RIKEN Center for Integrative Medical Sciences, Yokohama, Japan
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).