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  • ChIP-Seq from Limited Starting Material of K562 Cells and Drosophila Neuroblasts Using Tagmentation Assisted Fragmentation Approach.

ChIP-Seq from Limited Starting Material of K562 Cells and Drosophila Neuroblasts Using Tagmentation Assisted Fragmentation Approach.

Bio-protocol (2021-03-04)
Junaid Akhtar, Piyush More, Steffen Albrecht
要旨

Chromatin immunoprecipitation is extensively used to investigate the epigenetic profile and transcription factor binding sites in the genome. However, when the starting material is limited, the conventional ChIP-Seq approach cannot be implemented. This protocol describes a method that can be used to generate the chromatin profiles from as low as 100 human or 1,000 Drosophila cells. The method employs tagmentation to fragment the chromatin with concomitant addition of sequencing adaptors. The method generates datasets with high signal to noise ratio and can be subjected to standard tools for ChIP-Seq analysis.

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Sigma-Aldrich
Triton X-100, Molecular Biology
Sigma-Aldrich
ホルムアルデヒド 溶液, Molecular Biology, 36.5-38% in H2O
Sigma-Aldrich
塩化ナトリウム, Molecular Biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
トリス(ヒドロキシメチル)アミノメタン, ACS reagent, ≥99.8%
Roche
グリコーゲン, from mussels
Sigma-Aldrich
10 mMトリス、pH 8.0、1 mM EDTAで飽和した25:24:1のフェノール:クロロホルム:イソアミルアルコール, Molecular Biology
Sigma-Aldrich
グリシン, BioUltra, Molecular Biology, ≥99.0% (NT)
Sigma-Aldrich
N,N-ジメチルホルムアミド, Molecular Biology, ≥99%
Sigma-Aldrich
ウシ血清アルブミン ウシ血清由来, lyophilized powder, protease, essentially free, ≥98% (agarose gel electrophoresis)
Sigma-Aldrich
リボ核酸, トランスファー パン酵母(S. cerevisiae種)由来, Type X-SA, lyophilized powder
USP
コラゲナーゼI, United States Pharmacopeia (USP) Reference Standard