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Team:KAIST Korea/Notebook Labnote/2012 10
From 2012.igem.org
2012 KAIST Korea
Mail : kaist.igem.2012@gmail.com
Twitter : twitter.com/KAIST_iGEM_2012
Facebook : www.facebook.com/KAISTiGEM2012
Notebook : Labnote-October
Labnote
OctoberOctober 1st 2012
No Special Event!
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Back to the CalendarOctober 2nd 2012
No Special Event!
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October 3rd 2012
Flip FlopBxb1 double transformants vector miniprep & PCR check
Results
Discussions
Seemed to every colonies have appropriate vectors.
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October 4th 2012
No Special Event!
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October 5th 2012
No Special Event!
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October 6th 2012
No Special Event!
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October 7th 2012
No Special Event!
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October 8th 2012
No Special Event!
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October 9th 2012
No Special Event!
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October 10th 2012
Flip FlopBxb1_GTG_pBAD single cut check with different enzyme
Single cut with SnaBI
Results
No desired size(5.6kb) vector appeared.
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Flip Flop
Flip Flop
Flip Flop
Flip Flop
October 11th 2012
No Special Event!
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October 12th 2012
Flip FlopBxb1_GTG_pBAD colony #4~7 inoculation, miniprep and single cut check
Single cut with SnaBI
Single cut with SnaBI
Results
Cloning completed
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October 13th 2012
No Special Event!
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October 14th 2012
No Special Event!
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October 15th 2012
Flip FlopBBa_C0060, BBa_C0061, BBa_C0062 transformation into TOP10
Templates for Auto-regulated FlipFlop project.
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October 16th 2012
Flip FlopBBa_C0060, BBa_C0061, BBa_C0062, bFMO inoculation and enzyme cut check
BBa_C0060, BBa_C0061, BBa_C0062: from colony, EcoRI single cut
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- BBa_C0060: 2859bp
- BBa_C0061: 2688bp
- BBa_C0062: 2826bp
- bFMO: from cell stock, EcoRI single cut
October 17th 2012
Flip FlopTemplate preparation for OE PCR
Promoter construct: 210bp
Results
Promoter PCR failed
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Flip Flop
Flip Flop
Flip Flop
Flip Flop
Flip Flop
October 18th 2012
Flip FlopTemplate preparation for OE PCR
Results
Terminator PCR failed
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October 19th 2012
Flip FlopTemplate preparation for OE PCR
Terminator: 179bp
Results
→ Gel extracted
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October 20th 2012
Flip FlopTemplate preparation for OE PCR
Bxb1 Xis: 777bp
Results
bFMO PCR w/ various condtions
Bxb1_Xis-pGEM_B1 transfomation
Synthesized gene – Bxb1_Xis arrived and transformed into TOP10
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October 21st 2012
No Special Event!
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October 22nd 2012
Flip FlopOverlapping extension PCR
Results
→ Gel extracted
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October 23rd 2012
Flip FlopOverlapping extension PCR-Nested PCR
Results
Discussions
There was too little full construct nested. We think this is due to primer interfering by its LVA homology.
Overlapping extension PCR 2
Results
Discussions
Bands were more intense than those of first trial.
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October 24th 2012
No Special Event!
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October 25th 2012
No Special Event!
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October 26th 2012
No Special Event!
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October 27th 2012
No Special Event!
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October 28th 2012
No Special Event!
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October 29th 2012
No Special Event!
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October 30th 2012
No Special Event!
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October 31st 2012
No Special Event!
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