Minnesota/30 July 2008

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|'''[[Team:Minnesota/NotebookComparator|  Back to Notebook Home]]'''
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|'''[[Minnesota/29 July 2008|Go to Previous Day (July 29)]]'''|| width=158|'''[[Minnesota/31 July 2008|Go to Next Day (July 31)]]'''
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|''' 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.
[[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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|'''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:
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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.

Latest revision as of 19:51, 30 July 2008

Back to Notebook Home
Go to Previous Day (July 29)Go to Next Day (July 31)
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.
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.
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
BV + Lac/LAMBDA 5.0uL 4.0uL 2.0uL 25.0uL 4.0uL
NOTE: Used more Ligase (ligation enzyme) this time to attempt to ligate better.
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