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文献和实验He Tube Mating with MSCV recipient
the dilution with the OD closest to 0.1 (this is true for recipient AND donor cultures. You want an actively growing culture for optimal mating) 5. To set up tube mating, you want a 1:1 ratio of recipient:donor. Determine which dilution of the MSCV dilutions
Mix a 5-µL aliquot of the phosphopeptides affinity-bound to the IMAC media (from Step 10) with 5 µL of 50 mM NH4HCO3 (pH 7.8) and 1 unit of calf intestine alkaline phosphatase in a 0.7-mL microcentrifuge tube. Incubate the tube on a slowly rotating
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
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