Haynes Lab:Notebook/Engineering PC-TFs/2013/10/21: Difference between revisions
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== | ==October 21,2013== | ||
* | '''Miniprep liquid cultures''' | ||
*Add 100 μL of 7X Lysis Buffer (Blue) to 600 μL of E.coli culture in a 1.5 ml microcentrifuge tube. Mix by inverting tube 4-6times. | |||
*Add 350 μL of cold Neutralization Buffer (Yellow), mix thoroughly. Neutralization is complete when sample becomes yellow and precipitate has formed. | |||
*Centrifuge at 11,000-16,000 g for two minutes. | |||
*Transfer the supernatent into the Zymo-Spin Column. Place column in 2 ml collection tube and centrifuge for 15 seconds at top speed. Discard flow-through. | |||
*Add 200μL of Endo-Wash Buffer to the column. Centrifuge for 15 seconds at top speed. | |||
*Add 400μL of Zyppy Wash Buffer to the column. Centrifuge at top speed for 30 seconds. | |||
*Transfer column to 1.5 microcentrifuge tube and add 50μL of Zyppy Elution Buffer directly to column. Centrifuge for 1 minute at top speed to elute DNA. | |||
Revision as of 09:19, 25 October 2013
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October 21,2013Miniprep liquid cultures
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