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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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Triton X-100, for molecular biology
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ホルムアルデヒド 溶液, for molecular biology, 36.5-38% in H2O
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塩化ナトリウム, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
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グリシン, BioUltra, for molecular biology, ≥99.0% (NT)
Roche
グリコーゲン, from mussels
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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, for molecular biology
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N,N-ジメチルホルムアミド, for molecular biology, ≥99%
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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
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ウシ血清アルブミン ウシ血清由来, lyophilized powder, protease, essentially free, ≥98% (agarose gel electrophoresis)
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トリス(ヒドロキシメチル)アミノメタン, ACS reagent, ≥99.8%
USP
コラゲナーゼI, United States Pharmacopeia (USP) Reference Standard