- 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
PMID33654745
ABSTRAKT
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
Phenol:Chloroform:Isoamyl Alcohol 25:24:1, Saturated with 10mM Tris, pH 8.0, 1mM EDTA, for molecular biology
Sigma-Aldrich
Sodium chloride, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
Ribonucleic acid, transfer from baker′s yeast (S. cerevisiae), Type X-SA, lyophilized powder