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Minnesota/22 July 2008
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| - | |'''1. Pick Colonies from plates made 07-21-2008'''Start cultures. | + | |'''1. Pick Colonies from plates made 07-21-2008''' Start cultures. |
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|'''2. Plasmid prep:''' Prep RFP, YFP, GFP, TetR promoter, Terminator. Follow QIA miniprep procedure --> 1hr long. | |'''2. Plasmid prep:''' Prep RFP, YFP, GFP, TetR promoter, Terminator. Follow QIA miniprep procedure --> 1hr long. | ||
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|'''3. Double digest:''' Follow Kat's DNA work procedure to perform Double digest on: LacI Promoter, p22 cII gene, RFP, YFP, BV (dual promoters, GFP:Term already d.dig.). Incubate digested products for 2 hours @37C. Heat inactivate digestion enzyme for 15 mins @ 65C water bath. | |'''3. Double digest:''' Follow Kat's DNA work procedure to perform Double digest on: LacI Promoter, p22 cII gene, RFP, YFP, BV (dual promoters, GFP:Term already d.dig.). Incubate digested products for 2 hours @37C. Heat inactivate digestion enzyme for 15 mins @ 65C water bath. | ||
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| - | |'''4. Vector Dephosphorylation:''' | + | |'''4. Vector Dephosphorylation:''' Same dephos. procedure used on GFP:Terminator, BV. After dephosphorylation, incubate @37C for 30 mins. Heat inactivate dephosphorylation enzyme for 15 mins in 65C water bath. |
| + | |- | ||
| + | |'''5. Ligation Reactions:''' | ||
Revision as of 20:50, 21 July 2008
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| 1. Pick Colonies from plates made 07-21-2008 Start cultures. |
| 2. Plasmid prep: Prep RFP, YFP, GFP, TetR promoter, Terminator. Follow QIA miniprep procedure --> 1hr long. |
| 3. Double digest: Follow Kat's DNA work procedure to perform Double digest on: LacI Promoter, p22 cII gene, RFP, YFP, BV (dual promoters, GFP:Term already d.dig.). Incubate digested products for 2 hours @37C. Heat inactivate digestion enzyme for 15 mins @ 65C water bath. |
| 4. Vector Dephosphorylation: Same dephos. procedure used on GFP:Terminator, BV. After dephosphorylation, incubate @37C for 30 mins. Heat inactivate dephosphorylation enzyme for 15 mins in 65C water bath. |
| 5. Ligation Reactions: |
