Shreffler:Notebook/retinoic acid trans signaling/2009/05/20: Difference between revisions
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( Note: This process is used for rapid purification of transfection grade plasmid DNA) | ( Note: This process is used for rapid purification of transfection grade plasmid DNA) | ||
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temperature for 5 min | temperature for 5 min | ||
4. | - During the incubation prepare the QIAfilter Cartrigde | ||
- Screw the cap onto the outlet nozzle of the QIAfilter Maxi Cartridge | |||
- Place the QIAfilter Cartridge into a convenient tube or QIArack | |||
4. Add 10 ml of chilled Buffer P3, mix throughly by vigorously inverting 4-6 times | |||
B) Bacterial lysate clearing | |||
5. Pour the lysate into the barrel of the QIAfilter Catridge, and incubate at room temperature (15-20 degrees C) for 10 min, do not insert the plunger! | |||
6. Equilibrate a HiSpeed Maxi Tip by applying 10 ml Buffer QBT and allow the column to empty by gravity flow | |||
7. Remove the cap from the QIAfilter Cartridge outlet | |||
Revision as of 12:29, 1 June 2009
 Retinoic Acid Trans Signaling
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Plasmid Purification: ( Note: This process is used for rapid purification of transfection grade plasmid DNA)
A) Bacterial culture, harvest, and lysis 1. Pellet 150 ml overnight LB culture at 6000 x g for 15 min at 4 degree C 2. Homogeneously resuspend the bacterial pellet in 10 ml Buffer P1 3. Add 10 ml Buffer P2, mis throughly by vigoorously inverting 4-6 times, and incubate at room temperature for 5 min - During the incubation prepare the QIAfilter Cartrigde - Screw the cap onto the outlet nozzle of the QIAfilter Maxi Cartridge - Place the QIAfilter Cartridge into a convenient tube or QIArack 4. Add 10 ml of chilled Buffer P3, mix throughly by vigorously inverting 4-6 times
5. Pour the lysate into the barrel of the QIAfilter Catridge, and incubate at room temperature (15-20 degrees C) for 10 min, do not insert the plunger! 6. Equilibrate a HiSpeed Maxi Tip by applying 10 ml Buffer QBT and allow the column to empty by gravity flow 7. Remove the cap from the QIAfilter Cartridge outlet
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