Author: Steven Henikoff; Jorja G. Henikoff
Title: Profiling the epigenome at home Document date: 2020_4_17
ID: mojf4l6b_3
Snippet: More recently, we substituted the Tn5 transposase for MNase in a modified CUT&RUN protocol, such that addition of Mg ++ results in a cut-and-paste "tagmentation" reaction, in which sequencing adapters are integrated around sites of antibody binding (Kaya-Okur et al., 2019) . In CUT&Tag, DNA purification is followed by PCR amplification, eliminating the end-polishing and ligation steps required for sequencing library preparation in CUT&RUN. Like C.....
Document: More recently, we substituted the Tn5 transposase for MNase in a modified CUT&RUN protocol, such that addition of Mg ++ results in a cut-and-paste "tagmentation" reaction, in which sequencing adapters are integrated around sites of antibody binding (Kaya-Okur et al., 2019) . In CUT&Tag, DNA purification is followed by PCR amplification, eliminating the end-polishing and ligation steps required for sequencing library preparation in CUT&RUN. Like CUT&RUN, CUT&Tag requires relatively little input material, and the low backgrounds permit low sequencing depths to sensitively map chromatin features. Because integrated pA-Tn5 is not released following the tagmentation reaction, CUT&Tag and related methods are suitable for single-cell profiling, in which all steps through tagmentation are performed in a single in situ reaction, after which single cells or nuclei are dispensed for barcoding PCR amplification.
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