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Mnase enzyme

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Micrococcal Nuclease (MNase) is a widely used enzyme for chromatin and nucleosome research. It is an endonuclease that non-specifically cleaves DNA between nucleosomes, generating mononucleosomes and oligonucleosomes. The enzyme is useful for studying chromatin structure and dynamics.

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Lab products found in correlation

Nebnext ultra 2 dna library prep kit, by New England Biolabs (2 mentions) Nuclear isolation buffer, by Merck Group (2 mentions) H3k27me3, by Active Motif (2 mentions) H3k9me3, by Abcam (2 mentions)

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MNase (118 citations)

2 protocols using mnase enzyme

1

MNase Native ChIP-seq of Frozen Tissue Nuclei

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Purified nuclei from frozen tissue sections were subjected to MNase native ChIP-seq following the ULI NChIP procedure, as published elsewhere65 (link). Briefly, nuclei were suspended in Nuclear Isolation Buffer (Sigma) supplemented with 1% TritonX 100, 1% Deoxycholate and 1X complete protease inhibitor. Chromatin was digested by MNase enzyme (NEB, 1:10 diluted) at 21°C for 7.5 min, and further diluted in Complete Immunoprecipitation Buffer, with 1X complete protease inhibitor. 2 μl ChIP-grade H3K27me3 (Active motif, Cat#39155) or H3K9me3 (Abcam, Cat#ab8898) antibody was incubated with the digested chromatin overnight at 4°C. DNA was then purified using protease K digestion followed by phenol-chloroform extraction. ChIP-seq sequencing libraries were prepared using NEBNext Ultra II DNA Library Prep Kit (NEB, Cat#E7645L).
Guo H., Vuille J.A., Wittner B.S., Lachtara E.M., Hou Y., Lin M., Zhao T., Raman A.T., Russell H.C., Reeves B.A., Pleskow H.M., Wu C.L., Gnirke A., Meissner A., Efstathiou J.A., Lee R.J., Toner M., Aryee M.J., Lawrence M.S., Miyamoto D.T., Maheswaran S, & Haber D.A. (2023). DNA hypomethylation silences antitumor immune genes in early prostate cancer and CTCs. Cell, 186(13), 2765-2782.e28.
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2

MNase Native ChIP-seq of Frozen Tissue Nuclei

Check if the same lab product or an alternative is used in the 5 most similar protocols
Purified nuclei from frozen tissue sections were subjected to MNase native ChIP-seq following the ULI NChIP procedure, as published elsewhere65 (link). Briefly, nuclei were suspended in Nuclear Isolation Buffer (Sigma) supplemented with 1% TritonX 100, 1% Deoxycholate and 1X complete protease inhibitor. Chromatin was digested by MNase enzyme (NEB, 1:10 diluted) at 21°C for 7.5 min, and further diluted in Complete Immunoprecipitation Buffer, with 1X complete protease inhibitor. 2 μl ChIP-grade H3K27me3 (Active motif, Cat#39155) or H3K9me3 (Abcam, Cat#ab8898) antibody was incubated with the digested chromatin overnight at 4°C. DNA was then purified using protease K digestion followed by phenol-chloroform extraction. ChIP-seq sequencing libraries were prepared using NEBNext Ultra II DNA Library Prep Kit (NEB, Cat#E7645L).
Guo H., Vuille J.A., Wittner B.S., Lachtara E.M., Hou Y., Lin M., Zhao T., Raman A.T., Russell H.C., Reeves B.A., Pleskow H.M., Wu C.L., Gnirke A., Meissner A., Efstathiou J.A., Lee R.J., Toner M., Aryee M.J., Lawrence M.S., Miyamoto D.T., Maheswaran S, & Haber D.A. (2023). DNA hypomethylation silences antitumor immune genes in early prostate cancer and CTCs. Cell, 186(13), 2765-2782.e28.
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