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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
摘要

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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氯化钠, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
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甲醛 溶液, for molecular biology, 36.5-38% in H2O
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甘氨酸, BioUltra, for molecular biology, ≥99.0% (NT)
Roche
糖原, from mussels
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苯酚:氯仿:异戊醇25:24:1,10mM Tris饱和溶液,pH8.0,1mM EDTA, for molecular biology
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N,N-二甲基甲酰胺, for molecular biology, ≥99%
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三(羟甲基)氨基甲烷, ACS reagent, ≥99.8%
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核糖核酸,转移 来源于面包酵母(酿酒酵母), Type X-SA, lyophilized powder
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胶原酶I, United States Pharmacopeia (USP) Reference Standard