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文献和实验Dialysis (using dialysis tubing)
, deionized water, or 100% ethanol. Keep the tubing completely submersed. Uncleaned membranes can be stored at room temperature. 4.Do not autoclave . Heating over 80℃ changes the membrane characteristics (drop permeability). 5.To prevent
Preparation of Microinjection Buffer
Buffer composition is 10 mM Tris-HCl, pH 7.5, 0.1 mM EDTA, 30 microM spermine, 70 microM spermidine, 100 mM NaCl. This buffer is more likely to produce transgenic mice with intact, unfragmented DNA molecules. See the following references: Schedl
CCMB buffer to make chemical competent cell
on ice 20 minutes Centrifuge again at 4C and resuspend in 20 ml of ice cold CCMB80 buffer. Incubate on ice for 20 minutes Aliquot to chilled screw top 2 ml vials or 50 μl into chilled microtiter plates Store at -80C indefinitely. Flash freezing
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