We present a detailed step-wise protocol for reliable genome-wide profiling of histone modifications using CUT&RUN that requires only low numbers of Plasmodium falciparum blood stages. We recently adapted a CUT&RUN… Click to show full abstract
We present a detailed step-wise protocol for reliable genome-wide profiling of histone modifications using CUT&RUN that requires only low numbers of Plasmodium falciparum blood stages. We recently adapted a CUT&RUN protocol for genome-wide profiling of chromatin modifications in the human malaria parasite Plasmodium. Using the step-by-step protocol described below, we were able to generate high-quality profiles of multiple histone modifications using only a small fraction of the cells required for ChIP-seq. Using antibodies against two commonly profiled histone modifications, H3K4me3 and H3K9me3, we show here that CUT&RUN profiling is highly reproducible and closely recapitulates previously published ChIP-seq-based abundance profiles of histone marks. Finally, we show that CUT&RUN requires substantially lower sequencing coverage for accurate profiling compared with ChIP-seq.
               
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