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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

No Special Event!

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September 5^th 2012

No Special Event!

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September 6^th 2012

No Special Event!

Back to the Calendar

September 7^th 2012

No Special Event!

Back to the Calendar

September 8^th 2012

No Special Event!

Back to the Calendar

September 9^th 2012

No Special Event!

Back to the Calendar

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.
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September 11^th 2012

No Special Event!

Back to the Calendar

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

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September 13^th 2012

No Special Event!

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September 14^th 2012

No Special Event!

Back to the Calendar

September 15^th 2012

　Flip Flop

BBa_C0060, BBa_C0061, BBa_C0062 transformation into TOP10

Templates for Auto-regulated FlipFlop project.

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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

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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

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September 21^st 2012

No Special Event!

Back to the Calendar

September 22^nd 2012

　Flip Flop

Overlapping extension PCR

Results

→ Gel extracted

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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.

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September 24^th 2012

　Flip Flop

Overlapping extension PCR Gel Extraction

Results

P1, PI construct

Results

Full fragment for pAuto is overlapped. The right size(3794bp) fragments are gel extracted.

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September 25^th 2012

No Special Event!

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September 26^th 2012

　Flip Flop

Full pAuto and pAutoSimple Overlapping PCR

Results

Discussion

The band intensity is too weak.

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September 27^th 2012

No Special Event!

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September 28^th 2012

No Special Event!

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September 29^th 2012

No Special Event!

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September 30^th 2012

No Special Event!

Back to the Calendar

Kaist Footer

@@ Line 700: / Line 700: @@
 		<section id="26">
 			<div class="date">September 26<sup>th</sup> 2012</div></br>
-<div id="content_note" >
+				<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>
-				<span id="little">No Special Event!</span>
+			<div class="note-title">Full pAuto and pAutoSimple Overlapping PCR</div>
-				</br>
+			<div id="content_note" >
+				<b>Results</b></br></br>
+				<div align="center"><img id="figure" alt="0818Fig1" src="https://static.igem.org/mediawiki/2012/2/21/120926-1.PNG"></img></div>
+				<div style="clear:both;"></div>
+				</br></br>
+				<b>Discussion</b></br></br>
+				<span id="little">The band intensity is too weak.</span>
 			</div>
 			<a id="back_to_the_calender" style="font-size:12px;color:#111;text-decoration:none;float:right;margin:20px 0px 0px 5px;" href="https://2012.igem.org/wiki/index.php?title=Team:KAIST_Korea/Notebook_Labnote"><img src="https://static.igem.org/mediawiki/2012/e/e3/KAIST_calendar_icon.png" style="width:60px;height:60px;display:none;"/>Back to the Calendar</a>

Team:KAIST Korea/Notebook Labnote/2012 9

From 2012.igem.org

Revision as of 22:06, 26 October 2012

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