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Minnesota/24 July 2008
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|'''4. Vector Dephosphorylation:''' After the digested products' enzyme has been heat inactivated, we dephosphorylated the base vector so the cut ends wouldn't religate. Follow the table below: | |'''4. Vector Dephosphorylation:''' After the digested products' enzyme has been heat inactivated, we dephosphorylated the base vector so the cut ends wouldn't religate. Follow the table below: | ||
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Revision as of 16:24, 24 July 2008
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| 1. Base Vector Plasmid Maxi Prep Base Vector: Grew another culture of different base vector colonies, so now need to plasmid prep the base vector so the DNA/plasmid will be extracted from the rest of the cell. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 2. Spectrophotometry: Excellent results from spec'ing new base vector plasmid. Very high concentration of DNA. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 3. Base Vector & Promoter Double Digest: 50uL Reaction Mixture. Digest incubation for 2 hours @ 37C. Heat inactivate enzyme for 15 minutes @ 65C in a water bath. Follow the table below for pre-incubation steps:
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| 4. Vector Dephosphorylation: After the digested products' enzyme has been heat inactivated, we dephosphorylated the base vector so the cut ends wouldn't religate. Follow the table below:
|
