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.
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Engineering PC-TFs
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