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Minnesota/30 July 2008
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| - | |'''Gel electrophoresis''' was performed to separate digested LacI/Lambda cI dually-repressed promoter from the plasmid backbone. Lanes (L -> R) contain: 1kb ladder, two lanes of promoter digest, and a 100 bp ladder. | + | |''' Paper and Powerpoint for BSI''' - Work on them and begin practicing for presentation. |
| + | |- | ||
| + | |''' 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. | ||
| + | |- | ||
| + | |'''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. | ||
[[Image:Gel7.30.08.jpg|500px||center|thumb|Electrophoretic gel run on 7.30.08]] | [[Image:Gel7.30.08.jpg|500px||center|thumb|Electrophoretic gel run on 7.30.08]] | ||
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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: | |'''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: | ||
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| + | {|border="1" align="left" | ||
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| + | !| 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. | ||
Revision as of 19:50, 30 July 2008
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| Paper and Powerpoint for BSI - Work on them and begin practicing for presentation. | ||||||||||||
| 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. | ||||||||||||
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.
 | ||||||||||||
| Gel Purification - purified the DNA bands from the gel to extract only DNA and purify out cell debris. | ||||||||||||
| 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:
| ||||||||||||
| NOTE: Used more Ligase (ligation enzyme) this time to attempt to ligate better. |
