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Terri Bryson, Steven Henikoff 2019. 3XFlag-pATn5 Protein Purification and MEDS-loading (5x scale, 2L volume). protocols.io dx.doi.org/10.17504/protocols.io.8yrhxv6Copy Citation Copied
URL: https://dx.doi.org/10.17504/protocols.io.8yrhxv6
Authors: Terri Bryson, Steven Henikoff
Summary: Here we describe the method used to purify and load pATn5 for CUT&Tag, https://www.protocols.io/view/bench-top-cut-amp-tag-z6hf9b6/abstract. This protocol was modeled after that in Picelli et al (Picelli et al., 2014) using the pTXB1-rbs_3XFlag-pATn5-FL plasmid available from Addgene: https://www.addgene.org/124601/. It yields, as written, used at 1:200 dilution in CUT&Tag reactions, and for future use, both of which should be stored at -20C and can easily be scaled down.
Associated Publications: Henikoff S, Henikoff JG, Kaya-Okur HS, Ahmad K, Efficient chromatin accessibility mapping in situ by nucleosome-tethered tagmentation. eLife doi: 10.7554/eLife.63274
Affiliations: Fred Hutchinson Cancer Research Center, HHMI, Fred Hutchinson Cancer Research Center
External URL: https://www.nature.com/articles/s41467-019-09982-5
Version: 1
Publication Date: 2019
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Source: Protocols.io
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