MNase Digestion and Genomic DNA Purification

MNase assays were performed following the protocol as described previously 3 (link), 39 (link). Briefly, isolated nuclei were digested with 10 U/100 µL MNase in digestion buffer (15 mM Tris-HCl, pH7.4, 15 mM NaCl, 60 mM KCl, 1 mM CaCl₂, 0.25 M sucrose, and 0.5 mM DTT) at 37 °C for 10 min. The digested genomic DNA was carefully purified using a DNA purification kit (Axygen, #AP-PCR-250) and subjected to 1.2% agarose gel electrophoresis.

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Zhang F.L., Yang S.Y., Liao L., Zhang T.M., Zhang Y.L., Hu S.Y., Deng L., Huang M.Y., Andriani L., Ma X.Y., Shao Z.M, & Li D.Q. (2023). Dynamic SUMOylation of MORC2 orchestrates chromatin remodelling and DNA repair in response to DNA damage and drives chemoresistance in breast cancer. Theranostics, 13(3), 973-990.

Publication 2023

#AP-PCR-250

A dna Agarose gel electrophoresis Assays Buffer Digestion Genomic Nacl Nuclei Sucrose Tris

Corresponding Organization : Shanghai Medical College of Fudan University

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Variable analysis

independent variables

MNase concentration (10 U/100 μL)

dependent variables

Digested genomic DNA

control variables

Digestion buffer (15 mM Tris-HCl, pH7.4, 15 mM NaCl, 60 mM KCl, 1 mM CaCl2, 0.25 M sucrose, and 0.5 mM DTT)
Digestion temperature (37 °C)
Digestion time (10 min)

controls

Positive control: Not specified
Negative control: Not specified

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