Team:PKU Beijing/Notebook/Protocol/Miniprep
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12. Add 30-50 ul elution buffer (EB) to elute the DNA. | 12. Add 30-50 ul elution buffer (EB) to elute the DNA. | ||
- | '''Notes:''' | + | '''Notes:''' <br> |
1. Typical yield of high-copy-number plasmids, such as PSB1AK3, prepared by this method is about 0.2-0.3 ug of DNA per ul of original bacterial culture, and 0.1 ug of DNA per ul for low-copy-number plasmids such as PSB3T5.<br> | 1. Typical yield of high-copy-number plasmids, such as PSB1AK3, prepared by this method is about 0.2-0.3 ug of DNA per ul of original bacterial culture, and 0.1 ug of DNA per ul for low-copy-number plasmids such as PSB3T5.<br> | ||
2. To analyze the DNA by cleavage with restriction enzyme(s) remove 2 µl of the DNA solution and add it to fresh microfuge tube that contains 5 µl of water. Add 1 µl of the appropriate 10 x restriction enzyme(s). Incubate the reaction for 2 hr at the appropriate temperature. Store the remainder of the DNA preparation at -20 degree. Analyze the DNA fragments in the restriction digest by gel electrophoresis. <br> | 2. To analyze the DNA by cleavage with restriction enzyme(s) remove 2 µl of the DNA solution and add it to fresh microfuge tube that contains 5 µl of water. Add 1 µl of the appropriate 10 x restriction enzyme(s). Incubate the reaction for 2 hr at the appropriate temperature. Store the remainder of the DNA preparation at -20 degree. Analyze the DNA fragments in the restriction digest by gel electrophoresis. <br> |
Revision as of 07:55, 7 October 2009