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文献和实验Single tube confirmation PCR protocol
containing 60 U/ml of Zymolyase (This solution is generated by resuspending 6 mg of 100T Zymolyase in 10 mls of water. Ordering information: Seikagaku Corp., #120493-1). - Sterile yellow tips work great for this task. - Repeat this step
Purification of Plasmid from 50 ml-culture
of cold 70% ethanol. Discard the fluid. 14. Add to the pellet 400 ul of TE containing DNase-free RNase A (20 ug/ml). Incubate the tube for 30 min at 37 C. 15. After 30 min, carefully check the content of the tube. If nucleic acid pellet
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
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