Apr 17, 2020

Profiling the epigenome at home

BioRxiv : the Preprint Server for Biology
Jorja G. Henikoff, Steven Henikoff

Abstract

We recently described CUT&Tag, a general strategy for epigenomic profiing in which antibody-tethered Tn5 transposase integrates DNA sequencing adapters at sites of specific chromatin protein binding or histone modification in intact cells or nuclei. Here we introduce a simplified CUT&Tag method that can be performed at home to help ameliorate the interruption of bench research caused by COVID-19 physical distancing requirements. All steps beginning with frozen nuclei are performed in single PCR tubes through to barcoded library amplication and clean-up, ready for pooling and DNA sequencing. Our CUT&Tag@home protocol has minimal equipment, reagent and supply needs and does not require handling of toxic or biologically active materials. We show that data quality and reproducibility for samples down to ~100 nuclei compare favorably to datasets produced using lab-based CUT&Tag and other chromatin profiling methods. We use CUT&Tag@home with antibodies to trimethylated histone H3 lysine-4, -36, -27 and -9 to comprehensively profile the epigenome of human K562 cells, consisting respectively of active gene regulatory elements, transcribed gene bodies, developmentally silenced domains and constitutively silenced parasitic elements.

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Environment
Mutation Abnormality
Population Group
Biological Evolution
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