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- Beckman Coulter贝克曼库尔特
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- 2025年07月16日
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文献和实验Single tube confirmation PCR protocol
50 µl total volume *10 x Taq buffer contains: 100 mM Tris-HCl (pH 8.4), 500 mM KCl, 15 mM MgCl2. *The final concentrations are shown in parentheses. *The Taq Polymerase should be added last and PCR mixture should be kept on ice
Single tube confirmation PCR protocol
for two to three days. *This step reduces background by eliminating aborted transformants. 2. Zymolyase treatment - Transfer a small portion of a well-isolated colony (~1 mm) into 50 µl solution containing 60 U/ml of Zymolyase (This solution is generated
ES CELL DNA EXTRACTION: TUBE ME
off supernatant as in step 16. 19. Store the open tube on the bench at TRm until the last traces of fluid have evaporated. (i.e.: Air dry). 20. Dissolve the DNA in 30µl (or 50 µl, and use half of sample) of 10mM Tris pH 8.5 (or TE). (pipet
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