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文献和实验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
Purification of Plasmid from 50 ml-culture
of the tube with 500 ul 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
Single tube confirmation PCR protocol
(200mg/l). - Incubate at 30°C 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
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