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文献和实验CCMB buffer to make chemical competent cell
TOP10 chemically competent cells ( modified from http://openwetware.org/wiki/TOP10_chemically_competent_cells#CCMB80_buffer) I also use this protocol to make Ecoli compent cell. It is pretty good. You can start from the fresh clone on plate
Mitochondrial DNA Isolation from Somatic Embryogenic Cell Cultures of Larix (Excerpt from Thesis).
at 3000g for 10 minutes to separate cellular debris, nuclei and proplastids from the mitochondria. Mitochondria are pelleted at 10000g for 20 minutes, resuspended in DNase buffer (0.4M mannitol, 20mM HEPES, pH 7.5, 10mM magnesium chloride) and treated
ES / MEF cell culture and electroporation of targeting construct
the final volume up to 15 ml). Take this tube to the blood bank and irradiate (3000 rads). These inactivated MEFs will be used to make two 10cm dishes of MEF feeder layers for your ES cell thaw on Sunday. Using a hemacytometer, count the MEFs before adding
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