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Minnesota/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. | |'''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. | ||
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| - | |'''2. Spectrophotometry:''' Excellent results from spec'ing new base vector plasmid. Very high concentration of DNA. | + | |'''2. Spectrophotometry:''' Excellent results from spec'ing new base vector plasmid. Very high concentration of DNA, which was 430 ng/uL. |
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|'''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: | |'''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: | ||
Revision as of 16:30, 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, which was 430 ng/uL. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 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:
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