Author: Steven Henikoff; Jorja G. Henikoff
Title: Profiling the epigenome at home Document date: 2020_4_17
ID: mojf4l6b_2
Snippet: For example, in Cleavage Under Targets & Release Using Nuclease (CUT&RUN) (Skene and Henikoff, 2017) , which is based on Laemmli's Chromatin Immunocleavage (ChIC) method (Schmid et al., 2004) , a fusion protein between Micrococcal Nuclease (MNase) and Protein A binds sites of antibody binding in nuclei or permeabilized cells bound to magnetic beads. Activation of MNase with Ca ++ results in targeted cleavage releasing the antibody-bound fragment .....
Document: For example, in Cleavage Under Targets & Release Using Nuclease (CUT&RUN) (Skene and Henikoff, 2017) , which is based on Laemmli's Chromatin Immunocleavage (ChIC) method (Schmid et al., 2004) , a fusion protein between Micrococcal Nuclease (MNase) and Protein A binds sites of antibody binding in nuclei or permeabilized cells bound to magnetic beads. Activation of MNase with Ca ++ results in targeted cleavage releasing the antibody-bound fragment into the supernatant for paired-end DNA sequencing. The low backgrounds resulting from antibody-tethered cleavage and release reduce cell numbers and sequencing depths required to map chromatin features relative to ChIP-seq, and CUT&RUN is now becoming a standard tool for mapping transcription factors and other chromatin features (Hainer et al., 2019; Liu et al., 2018; Oomen et al., 2019; Roth et al., 2018) .
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