微球菌核酸酶

微球菌核酸酶，Micrococcal Nuclease，可裂解单链和双链 DNA 和 RNA。更多视频请关注视频号【艾维缔】。哔哩哔哩【IVDSHOW】。抖音【军哥聊表观】。

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微球菌核酸酶

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酶

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D5220-1

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微球菌核酸酶可裂解单链和双链 DNA 和 RNA。用微球菌核酸酶完全消化后，可得到带有 3′-磷酸的单核和寡核苷酸。

Concentration	0.1 U/µl
Enzyme Commission Number	(E.C. 3.1.31.1)
Enzyme Inactivation	EDTA or EGTA in molar excess of CaCl2.
Storage	Store at -20°C for up to 12 months. Avoid repeated freeze/thawing. Prolonged storage should be ≤ -70 °C.
Unit Definition	One unit (U) will produce 1.0 µmole of acid soluble polynucleotides from native DNA per min at pH 8.8 at 37°C, based on EM/260 = 10,000 for the mixed nucleotides.

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微球菌核酸酶
D5220-1 10U/100ul -20°C
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Micrococcal nuclease digestion was performed on embryonic stem cells using the EZ Nucleosomal DNA Prep Kit and paired-end libraries were generated for sequencing. Research showed heterogeneous chromatin organization around transcription start sites as well as unique nucleosome profiles associated with epigenetic patterns that can identify pluripotency.

Yazdi PG, et. al. (2015). Nucleosome Organization in Human Embryonic Stem Cells. PLoS ONE. 10(8).

The EZ Nucleosomal DNA Prep Kit was used by researchers to isolate nucleosomal DNA from 293FT cells prior to performing Chromatin Accessibility Real Time PCR (CHART-PCR) to investigate the IL-2 promoter chromatin architecture in the presence of TATA-box binding protein (TBP-TALE) and VP64-TALE activators in non-immune cells. They found that the combination of AD’CF TALEs but not empty vector control increased DNase I hypersensitivity across the IL-2 promoter.

Anthony K et al. (2014) Activation of Silenced Cytokine Gene Promoters by the Synergistic Effect of TBP-TALE and VP64-TALE Activators. PLoS One. 2014 Apr 22;9(4):e95790.

Researchers used the EZ Nucleosomal DNA Prep Kit from Zymo Research to isolate nuclei, digest chromatin, and purify nucleosomal DNA from Hep3B cells or SW13 cells that were transfected with either an empty vector or vectors expressing WT BRG1 or ATPase-dead BRG1. Purified nucleosomal DNA was used for SYBR Green–based qPCR to determine nucleosome positioning on the CA9 and LDHA promoters.

Sena JA et al. (2013) BRG1 and BRM chromatin remodeling complexes regulate the hypoxia response by acting as a co-activator for a subset of HIF target genes. Mol Cell Biol.

An MNase digestion assay was performed using the EZ Nucleosomal DNA Prep Kit to study the kinetics of micrococcal nuclease digestion in wild type and LAP2α deficient HeLa cells.

Zhang S et al. (2013) High Mobility Group Protein N5 (HMGN5) and Lamina Associated Polypeptide 2 alpha (LAP2α) Interact and Reciprocally Affect Their Genome-wide Chromatin Organization. J Biol Chem.

Mononucleosomal DNA from p16-methylated AGS and p16-unmethylated MGC803 cell lines was isolated using the EZ Nucleosomal DNA Prep Kit. The isolated mononucleosomal DNA was used to study the chromatin accessibility across the p16 promoter by quantitative PCR.

Lu ZM, Zhou J, Wang X, Guan Z, Bai H, Liu ZJ, Su N, Pan K, Ji J, Deng D. (2012) Nucleosomes correlate with in vivo progression pattern of de novo methylation of p16 CpG islands in human gastric carcinogenesis. PLoS One. 7(4):e35928.

The EZ Nucleosomal DNA Prep Kit from Zymo Research was used to isolate nuclei from mouse pituitary tissue and B cells. The purified DNA was further isolated into mononucleosome core fragments and sequenced for nucleosome mapping studies of the human growth hormone gene.

Yoo EJ et al. (2015) Autonomous actions of the human growth hormone long-range enhancer. Nucleic Acids Res. 43(4):2091-101. doi: 10.1093/nar/gkv093.

Nucleosomal DNA was isolated from mouse splenocytes using the EZ Nucleosomal DNA Prep Kit from Zymo Research. Nucleosomal DNA was used to demonstrate the effect of topo I on binding of anti-dsDNA antibodies. Surprisingly, topo I increased binding of these antibodies in the plasma of lupus-prone mice. This data, combined with additional findings, suggests a possible therapeutic target in topo I for Lupus patients.

Frese-Schaper M et al. (2014) Low‐Dose Irinotecan Improves Advanced Lupus Nephritis in Mice Potentially by Changing DNA Relaxation and Anti–Double‐Stranded DNA Binding. Arthritis Rheumatol. 66(8):2259-69. doi: 10.1002/art.38665.

Researchers from the University of Rochester performed micrococcal nuclease digestion of DNA using the EZ Nucleosomal DNA Prep Kit from Zymo Research. The authors showed that SIRT6 KO mouse embryonic fibroblasts had disturbed higher order chromatin structures. Further studies demonstrated that L1 loci were particularly sensitive to the micrococcal nuclease treatment and that SIRT6 is required for packaging L1 loci into transcriptionally repressive heterochromatin.

Van Meter M et al. (2014) SIRT6 represses LINE1 retrotransposons by ribosylating KAP1 but this repression fails with stress and age. Nat Commun. 5:5011. doi: 10.1038/ncomms6011.