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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
ZUSAMMENFASSUNG

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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Formaldehyd -Lösung, for molecular biology, 36.5-38% in H2O
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Natriumchlorid, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
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Glycin, BioUltra, for molecular biology, ≥99.0% (NT)
Roche
Glycogen, from mussels
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N,N-Dimethylformamid, for molecular biology, ≥99%
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Phenol/Chloroform/Isoamylalkohol (25:24:1), gesättigt mit 10 mM Tris, pH 8,0, 1 mM EDTA, for molecular biology
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Ribonukleinsäure, Transfer aus Backhefe (S. cerevisiae), Type X-SA, lyophilized powder
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Tris(hydroxymethyl)aminomethan, ACS reagent, ≥99.8%
USP
Kollagenase I, United States Pharmacopeia (USP) Reference Standard