Cell nucleus extraction method
A technology of cell nuclei and cardiomyocytes, applied in the field of biomedicine
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[0047] Extract nuclei as follows:
[0048] Step 1. Obtaining isolated canine cardiomyocytes, including: preparing oxygenated compound buffer solution and related instruments and perfusion devices; routinely processing experimental animals to obtain isolated experimental animal hearts; perfusion and digestion of isolated experimental animal hearts, obtaining cardiomyocytes;
[0049] Step 2, subcellular fractionation, which includes: homogenization of isolated experimental animal myocardial tissue and cardiomyocytes; differential centrifugation of tissue and cell homogenate to obtain separated crude nuclei, membranes and cytoplasm; discontinuous sucrose density gradient centrifugation After centrifugation after obtaining the coarse nuclear precipitation layer, the separated nucleus was obtained and showed an elongated almond shape or a round shape, and was positive for DNA staining; all sub-steps in step 2 were carried out on ice or at 4 degrees.
[0050] More specifically, inc...
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