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文献和实验Protein Expression and Purification of MBP-T antigen (131-259)
thrombin (200U for 6 L prep.)and dialyze sample against buffer CO/N in cold room.Make sure the cleavage is complete.If it is not complete let the reaction go at room temperature with another addition of 100μl of thrombin. xvi.Pass the sample over pre
the following DNA dilution, and aliquot 35 μl into ten 1.5 ml microcentrifuge tubes: DNA 100 μg 10X TM buffer 35 μlsterile ddH2 O q.s. Final Volume 350 μl 2. To determine the optimal sonication conditions, sonicate the DNA samples in five of the tubes
as before, decant and resuspend the competent cell pellet in 100 μl (1/10 volume) of calcium chloride. 6. Transfer each 100 μl sample of competent cells to chilled 12 x 75 mm Falcon tubes which contain 3 to 5 μl of DNA sample (about 2 ng/ul to 20 ng/ul
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