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文献和实验Construction of BAC Libraries：Megabase DNA Isolation
: Triton X-100 is mixed with 1 x HB without b-mercaptoethanol to 20%. The solution is stored at 4°C. Wash Buffer (1 x HB plus 0.5% Triton X-100): Prepared by mixing 1 x HB without b-mercaptoethanol with Triton X-l00 and stored
or cosmid DNA of interest into a 12 X 75 mm Falcon tube containing 2 ml of LB media supplemented with the appropriate antibiotic (typically ampicillin at 100 ug/ml) and incubate at 37deg C 8-10 hours with shaking at 250 rpm. Transfer the culture
OUTLINE OF PROCEEDINGS USED IN TISSUE CULTURE TRANSFECTIONS
. Mix evenly and incubate for 24 hours. 3. Split the cells to 1.5 x 105 cells per 100mm culture dish. Use I dish per transfection and one for the negative control. Remember to freeze any leftover cells. Mix evenly and incubate for 24 hours.
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