Team:Newcastle/Minipreps
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{{Team:Newcastle/mainbanner}} | {{Team:Newcastle/mainbanner}} | ||
- | + | =Minipreps= | |
- | == | + | ==Procedures== |
+ | The Miniprep Kit is used to extract plasmid. | ||
+ | # Resuspend pelleted bacterial cells in 250 μl Buffer P1 and transfer to a microcentrifuge tube. | ||
+ | # Add 250 μl Buffer P2 and mix throughly by inverting the tube 4-6 times. | ||
+ | # Add 350 μl Buffer N3 and mix immediately and thoroughly by inverting the tube 4-6 times. | ||
+ | # Centrifuge for 10 min at 13,000 rpm in a table-top microcentrifuge. | ||
+ | # Apply the supernatant to the QIAprep spin column by decanting or pipetting. | ||
+ | # Centrifuge for 30-60s. Discard the flow-through. | ||
+ | # Recommended: Wash the QIAprep spin column by adding 0.5 ml Buffer PB and centrifuging for 30-60s. Discard the flow-through. | ||
+ | # Wash QIAprep spin column by adding 0.75 ml Buffer PE and centrifuging for 30-60s. | ||
+ | # Discard the flow-through, and centrifuge for an additional 1 min to remove residual wash buffer. | ||
+ | # To elute DNA, place the QIAprep column in a clean 1.5 ml microcentrifuge tube. Add 50 μl Buffer EB or water to the center of each QIAprep spin column, let stand for 1 min, and centrifuge for 1 min. | ||
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Revision as of 15:07, 28 July 2010
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Minipreps
Procedures
The Miniprep Kit is used to extract plasmid.
- Resuspend pelleted bacterial cells in 250 μl Buffer P1 and transfer to a microcentrifuge tube.
- Add 250 μl Buffer P2 and mix throughly by inverting the tube 4-6 times.
- Add 350 μl Buffer N3 and mix immediately and thoroughly by inverting the tube 4-6 times.
- Centrifuge for 10 min at 13,000 rpm in a table-top microcentrifuge.
- Apply the supernatant to the QIAprep spin column by decanting or pipetting.
- Centrifuge for 30-60s. Discard the flow-through.
- Recommended: Wash the QIAprep spin column by adding 0.5 ml Buffer PB and centrifuging for 30-60s. Discard the flow-through.
- Wash QIAprep spin column by adding 0.75 ml Buffer PE and centrifuging for 30-60s.
- Discard the flow-through, and centrifuge for an additional 1 min to remove residual wash buffer.
- To elute DNA, place the QIAprep column in a clean 1.5 ml microcentrifuge tube. Add 50 μl Buffer EB or water to the center of each QIAprep spin column, let stand for 1 min, and centrifuge for 1 min.