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KAIST Korea 2012 iGEM

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2012 KAIST Korea

Mail : kaist.igem.2012@gmail.com
Twitter : twitter.com/KAIST_iGEM_2012
Facebook : www.facebook.com/KAISTiGEM2012

Notebook : Labnote-September

Labnote

September

September 1^st 2012

　Flip Flop

Bxb1_GTG_pBAD cloning with remaining DNAs

Remaining Bxb1_GTG insert was cloned into pBADmycHisC vector. And electrotransformed into MG1655.

Back to the Calendar

September 2^nd 2012

　Flip Flop

Bxb1_ATG_pTrc, pBAD, Bxb1_GTG_pTrc miniprep for double transformation, Bxb1_GTG_pBAD colony inoculation and vector miniprep NcoI single cut check.

Results

Discussions

All the vectors showed right sizes. But Bxb1_GTG_pBAD colony #1 through 3 have undesired bands. They may be the supercoiled vector.

Bxb1_ATG_pTrc, pBAD, Bxb1_GTG_pTrc double transformation into pPoC and pPoCpi

Back to the Calendar

September 3^rd 2012

　Flip Flop

Bxb1 double transformants vector miniprep & PCR check

Results

Discussions

Seemed to every colonies have appropriate vectors.

Back to the Calendar

September 4^th 2012

September 5^th 2012

September 6^th 2012

September 7^th 2012

September 8^th 2012

September 9^th 2012

September 10^th 2012

　Flip Flop

Bxb1_GTG_pBAD single cut check with different enzyme

Single cut with SnaBI

Results

No desired size(5.6kb) vector appeared.
Back to the Calendar

September 11^th 2012

September 12^th 2012

　Flip Flop

Bxb1_GTG_pBAD colony #4~7 inoculation, miniprep and single cut check Single cut with SnaBI

Single cut with SnaBI

Results

Cloning completed

Back to the Calendar

September 13^th 2012

September 14^th 2012

September 15^th 2012

　Flip Flop

BBa_C0060, BBa_C0061, BBa_C0062 transformation into TOP10

Templates for Auto-regulated FlipFlop project.

Back to the Calendar

September 16^th 2012

　Flip Flop

BBa_C0060, BBa_C0061, BBa_C0062, bFMO inoculation and enzyme cut check

BBa_C0060, BBa_C0061, BBa_C0062: from colony, EcoRI single cut

BBa_C0060: 2859bp
BBa_C0061: 2688bp
BBa_C0062: 2826bp
bFMO: from cell stock, EcoRI single cut

Results

Back to the Calendar

September 17^th 2012

　Flip Flop

Template preparation for OE PCR

Promoter construct: 210bp
Results

Promoter PCR failed

Back to the Calendar

September 18^th 2012

　Flip Flop

Template preparation for OE PCR

Results

Terminator PCR failed

Back to the Calendar

September 19^th 2012

　Flip Flop

Template preparation for OE PCR

Terminator: 179bp

Results

→ Gel extracted

Back to the Calendar

September 20^th 2012

　Flip Flop

Template 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

Back to the Calendar

September 21^st 2012

September 22^nd 2012

　Flip Flop

Overlapping extension PCR

Results

→ Gel extracted

Back to the Calendar

September 23^rd 2012

　Flip Flop

Overlapping 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.

Back to the Calendar

September 24^th 2012

　Flip Flop

Overlapping extension PCR Gel Extraction

Results

Back to the Calendar

September 25^th 2012

September 26^th 2012

September 27^th 2012

September 28^th 2012

September 29^th 2012

September 30^th 2012

September 31^st 2012

Kaist Footer

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 			<div class="date">September 17<sup>th</sup> 2012</div></br>
-			<div id="linegray" align="center" style="font-size:15px; font-weight:bold; color:#888;"><img style="height:3px;width:269px;margin:0px 5px 0px 0px; "src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/><img stylsght:3px;width:269px; margin:0px 0px 0px 5px;"src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/></div></br>
+			<div id="linegray" align="center" style="font-size:15px; font-weight:bold; color:#888;"><img style="height:3px;width:269px;margin:0px 5px 0px 0px; "src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/><img style="width:40px; height:28px;float:bottom;"id="flipflop" src='https://static.igem.org/mediawiki/2012/2/22/KAIST_FlipFlop.png'/>　Flip Flop<img style="height:3px;width:269px; margin:0px 0px 0px 5px;"src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/></div></br>
 			<div class="note-title">Template preparation for OE PCR</div>
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 		<section id="18">
 			<div class="date">September 18<sup>th</sup> 2012</div></br>
-				<div id="linegray" align="center" style="font-size:15px; font-weight:bold; color:#888;"><img style="height:3px;width:269px;margin:0px 5px 0px 0px; "src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/><img stylsght:3px;width:269px; margin:0px 0px 0px 5px;"src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/></div></br>
+				<div id="linegray" align="center" style="font-size:15px; font-weight:bold; color:#888;"><img style="height:3px;width:269px;margin:0px 5px 0px 0px; "src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/><img style="width:40px; height:28px;float:bottom;"id="flipflop" src='https://static.igem.org/mediawiki/2012/2/22/KAIST_FlipFlop.png'/>　Flip Flop<img style="height:3px;width:269px; margin:0px 0px 0px 5px;"src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/></div></br>
 			<div class="note-title">Template preparation for OE PCR</div>
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 		<section id="19">
 			<div class="date">September 19<sup>th</sup> 2012</div></br>
-				<div id="linegray" align="center" style="font-size:15px; font-weight:bold; color:#888;"><img style="height:3px;width:269px;margin:0px 5px 0px 0px; "src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/><img stylsght:3px;width:269px; margin:0px 0px 0px 5px;"src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/></div></br>
+				<div id="linegray" align="center" style="font-size:15px; font-weight:bold; color:#888;"><img style="height:3px;width:269px;margin:0px 5px 0px 0px; "src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/><img style="width:40px; height:28px;float:bottom;"id="flipflop" src='https://static.igem.org/mediawiki/2012/2/22/KAIST_FlipFlop.png'/>　Flip Flop<img style="height:3px;width:269px; margin:0px 0px 0px 5px;"src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/></div></br>
 			<div class="note-title">Template preparation for OE PCR</div>
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 			<div class="date">September 20<sup>th</sup> 2012</div></br>
-			<div id="linegray" align="center" style="font-size:15px; font-weight:bold; color:#888;"><img style="height:3px;width:269px;margin:0px 5px 0px 0px; "src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/><img stylsght:3px;width:269px; margin:0px 0px 0px 5px;"src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/></div></br>
+			<div id="linegray" align="center" style="font-size:15px; font-weight:bold; color:#888;"><img style="height:3px;width:269px;margin:0px 5px 0px 0px; "src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/><img style="width:40px; height:28px;float:bottom;"id="flipflop" src='https://static.igem.org/mediawiki/2012/2/22/KAIST_FlipFlop.png'/>　Flip Flop<img style="height:3px;width:269px; margin:0px 0px 0px 5px;"src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/></div></br>
 			<div class="note-title">Template preparation for OE PCR</div>
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 			<div class="date">September 22<sup>nd</sup> 2012</div></br>
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 			<div class="note-title">Overlapping extension PCR</div>
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 			<div class="date">September 23<sup>rd</sup> 2012</div></br>
-				<div id="linegray" align="center" style="font-size:15px; font-weight:bold; color:#888;"><img style="height:3px;width:269px;margin:0px 5px 0px 0px; "src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/><img stylsght:3px;width:269px; margin:0px 0px 0px 5px;"src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/></div></br>
+				<div id="linegray" align="center" style="font-size:15px; font-weight:bold; color:#888;"><img style="height:3px;width:269px;margin:0px 5px 0px 0px; "src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/><img style="width:40px; height:28px;float:bottom;"id="flipflop" src='https://static.igem.org/mediawiki/2012/2/22/KAIST_FlipFlop.png'/>　Flip Flop<img style="height:3px;width:269px; margin:0px 0px 0px 5px;"src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/></div></br>
 			<div class="note-title">Overlapping extension PCR-Nested PCR</div>
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 			<div class="date">September 24<sup>th</sup> 2012</div></br>
-				<div id="linegray" align="center" style="font-size:15px; font-weight:bold; color:#888;"><img style="height:3px;width:269px;margin:0px 5px 0px 0px; "src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/><img stylsght:3px;width:269px; margin:0px 0px 0px 5px;"src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/></div></br>
+				<div id="linegray" align="center" style="font-size:15px; font-weight:bold; color:#888;"><img style="height:3px;width:269px;margin:0px 5px 0px 0px; "src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/><img style="width:40px; height:28px;float:bottom;"id="flipflop" src='https://static.igem.org/mediawiki/2012/2/22/KAIST_FlipFlop.png'/>　Flip Flop<img style="height:3px;width:269px; margin:0px 0px 0px 5px;"src="https://static.igem.org/mediawiki/2012/1/1b/KAIST_horizontal_gray.png"/></div></br>
 			<div class="note-title">Overlapping extension PCR Gel Extraction</div>
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-			</br><div align="center"><img style="width:670px; height:2px;" src="https://static.igem.org/mediawiki/2012/7/7f/Horizontal_line_kaist.png"></img></div></br>
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Team:KAIST Korea/Notebook Labnote/2012 9

From 2012.igem.org

Revision as of 00:55, 27 September 2012

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