Haynes Lab:Notebook/Engineering PC-TFs/2012/04/10: Difference between revisions
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== | ==4/11/12== | ||
* | Miniprep for flyPCD set | ||
*Pellet 1 - 5 mL bacterial overnight culture by centrifugation at >8000 rpm for 3 min at room temperature | |||
*Resuspend pelleted bacterial cells in 250 µl Buffer P1 and transfer to microcentrifuge tube | |||
*Add 250 µl Buffer P2 and mix by inverting tube 6 times until solution is clear. | |||
*Add 350 µl Buffer N3 and mix by inverting the tube 6 times. | |||
*Centrifuge for 10 minutes at 13,000 rpm | |||
*Apply the supernatant liquid to the QIAprep spin by decanting. Centrifuge for 30-60 second and discard the flow-through. | |||
*Wash the QIAprep column by adding .75 ml Buffer PE | |||
*Centrifuge for 60 seconds and discard the flow through. | |||
*Centrifuge for 1 min to remove residual wash buffer. | |||
*Place the QIAprep column in a clean 1.5 ml tube. To remove the DNA, add water to the center of the QIA spin columns, and centrifuge for 2 min. | |||
Revision as of 00:21, 11 April 2012
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4/11/12Miniprep for flyPCD set
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