| Paper and Powerpoint for BSI - Work on them and begin practicing for presentation.
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| Pick Colonies from plates made 07-29-08: One plate has BV:TetR/p22 dual promoters: RFP (all ligated together), and other plate has BV:TetR promoter: LAMBDAcIw/RBS from PCR reaction (all ligated together). After picking, place the picked colonies in 2mL LB culture tubes and allow to incubate @37C for approximately 8 hours.
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Gel electrophoresis was performed to separate digested LacI/Lambda cI dually-repressed promoter from the plasmid backbone. Gel was run on L/L from yesterday. Lanes (L -> R) contain: 1kb ladder, two lanes of promoter digest, and a 100 bp ladder.
 Electrophoretic gel run on 7.30.08 |
| Gel Purification - purified the DNA bands from the gel to extract only DNA and purify out cell debris.
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| Ligation Reaction - Ligated BV + LacI/LAMBDAcI together. After following ligation table, allow to incubate @ 16C for 45 minutes. Then heat inactivate enzyme in 65C water bath for 15 minutes. Follow the table below:
| Parts | H20 | 10X Buffer | Base Vector | Insert part DNA | T4 DNA ligase
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| BV + Lac/LAMBDA | 5.0uL | 4.0uL | 2.0uL | 25.0uL | 4.0uL
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| NOTE: Used more Ligase (ligation enzyme) this time to attempt to ligate better.
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