Team:KAIST Korea/Notebook Labnote/2012 10

From 2012.igem.org

(Difference between revisions)
Line 402: Line 402:
</div>
</div>
<a id="back_to_the_calender" style="font-size:12px;color:#111;text-decoration:none;float:right;margin:20px 0px 0px 5px;" href="http://2012.igem.org/wiki/index.php?title=Team:KAIST_Korea/Notebook_Labnote"><img src="http://2012.igem.org/wiki/images/e/e3/KAIST_calendar_icon.png" style="width:60px;height:60px;display:none;"/>Back to the Calendar</a>
<a id="back_to_the_calender" style="font-size:12px;color:#111;text-decoration:none;float:right;margin:20px 0px 0px 5px;" href="http://2012.igem.org/wiki/index.php?title=Team:KAIST_Korea/Notebook_Labnote"><img src="http://2012.igem.org/wiki/images/e/e3/KAIST_calendar_icon.png" style="width:60px;height:60px;display:none;"/>Back to the Calendar</a>
 +
</section>
</section>
<section id="18">
<section id="18">
<div class="date">October 18<sup>th</sup> 2012</div></br>
<div class="date">October 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="http://2012.igem.org/wiki/images/1/1b/KAIST_horizontal_gray.png"/><img style="width:40px; height:28px;float:bottom;"id="flipflop" src='http://2012.igem.org/wiki/images/2/22/KAIST_FlipFlop.png'/> Flip Flop<img style="height:3px;width:269px; margin:0px 0px 0px 5px;"src="http://2012.igem.org/wiki/images/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="http://2012.igem.org/wiki/images/1/1b/KAIST_horizontal_gray.png"/><img style="width:40px; height:28px;float:bottom;"id="flipflop" src='http://2012.igem.org/wiki/images/2/22/KAIST_FlipFlop.png'/>Flip Flop<img style="height:3px;width:269px; margin:0px 0px 0px 5px;"src="http://2012.igem.org/wiki/images/1/1b/KAIST_horizontal_gray.png"/></div></br>
<div class="note-title">Cell culture in 96-well plate for Indigo expression time optimization</div>
<div class="note-title">Cell culture in 96-well plate for Indigo expression time optimization</div>
Line 425: Line 426:
-
<span id="little"></br>We subtracted absorbance value of pure LB from value of samples and we set all samples to have same amount of cell using O.D value </br></br></span>
+
<span id="little"></br></br>We subtracted absorbance value of pure LB from value of samples and we set all samples to have same amount of cell using O.D value </br>
 +
Graph shown below is indigo intensity per cell. After 3 days incubation, indigo seems to be degraded and all the samples seem to have almost same indigo level.</br></br></span>
 +
 
 +
<div align="center"><img id="figure" alt="0818Fig1" src="http://2012.igem.org/wiki/images/f/fc/Tecan_indigo-1.PNG"></img></div>
 +
<div style="clear:both;"></div></br>
 +
 
 +
<span id="little"></br>We excluded 0hr data from the graph to see teh rising tendency clearly.</br></br></span>
 +
 
 +
<div align="center"><img id="figure" src="http://2012.igem.org/wiki/images/0/06/Tecan_indigo-2.PNG"></img></div>
 +
<div style="clear:both;"></div>
-
<b>Discussion</b></br></br>
+
<b></br>Discussion</b></br>
<span id="little"></br>This data shows that the cell successfully produces indigo. We can use this gene as a reporter of our module. However, we only checked the increase of indigo. It may decrease after prolonged culture. We should do additional experiments to get the peak in the graph and optimize the production time of indigo. </br></span>
<span id="little"></br>This data shows that the cell successfully produces indigo. We can use this gene as a reporter of our module. However, we only checked the increase of indigo. It may decrease after prolonged culture. We should do additional experiments to get the peak in the graph and optimize the production time of indigo. </br></span>
Line 433: Line 443:
</div>
</div>
-
<a id="back_to_the_calender" style="font-size:12px;color:#111;text-decoration:none;float:right;margin:20px 0px 0px 5px;" href="http://2012.igem.org/wiki/index.php?title=Team:KAIST_Korea/Notebook_Labnote"><img src="http://2012.igem.org/wiki/images/e/e3/KAIST_calendar_icon.png" style="width:60px;height:60px;display:none;"/>Back to the Calendar</a>
+
<a id="back_to_the_calender" style="font-size:12px;color:#111;text-decoration:none;float:right;margin:20px 0px 0px 5px;" href="http://2012.igem.org/wiki/index.php?title=Team:KAIST_Korea/Notebook_Labnote"><img src="http://2012.igem.org/wiki/images/e/e3/KAIST_calendar_icon.png" style="width:60px;height:60px;display:none;"/>Back to the Calendar</a>
-
</section>
+
</section>
<section id="19">
<section id="19">
<div class="date">October 19<sup>th</sup> 2012</div></br>
<div class="date">October 19<sup>th</sup> 2012</div></br>

Revision as of 01:51, 27 October 2012

KAIST Korea 2012 iGEM

Notebook : Labnote-October

Labnote

October

October 1st 2012

No Special Event!
Back to the Calendar
October 2nd 2012

 Flip Flop

bFMO template preparation with new primer
Results

0818Fig1

We have found that our bFMO sequence was wrong. So that we have ordered new primers with right sequence.
Back to the Calendar
October 3rd 2012

 Flip Flop

Visualization of inversion from GFP to RFP
Procedure


bfmo gene is constructed in the vector(pJ401, Km registance) and transformed into DH5α.
50λ cell from stock is inoculated in 5mL LB


Visualization of inversion from GFP to RFP
Procedure

We sampled the cell every 3hr. We induced one group of samples and un-induced another group of samples as a control. We took the picture of cell with confocal microscope at absorbance 500nm for GFP expression and 600nm for RFP expression.
Results

0818Fig1


Discussion

We obtained proper data. Un-induced sample doesn’t show measurable RFP expression throughout the culture. However, induced sample shows the increase of RFP expression. Increase of GFP level in induced sample is due to the stability of GFP and time delay before inversion.


Insert DNA preparation with OE pcr
Results

0818Fig1


Cloning
Results

0818Fig1
0818Fig1
Back to the Calendar
October 4th 2012

No Special Event!
Back to the Calendar
October 5th 2012

Regional Jamboree at HKUST!!!!
Back to the Calendar
October 6th 2012

Regional Jamboree at HKUST!!!!
Back to the Calendar
October 7th 2012

Regional Jamboree at HKUST!!!!
Back to the Calendar
October 8th 2012

No Special Event!
Back to the Calendar
October 9th 2012

No Special Event!
Back to the Calendar
October 10th 2012

No Special Event!
Back to the Calendar
October 11th 2012

No Special Event!
Back to the Calendar
October 12th 2012

No Special Event!
Back to the Calendar
October 13th 2012

No Special Event!
Back to the Calendar
October 14th 2012

No Special Event!
Back to the Calendar
October 15th 2012

No Special Event!
Back to the Calendar
October 16th 2012

No Special Event!
Back to the Calendar
October 17th 2012

No Special Event!
Back to the Calendar
October 18th 2012

Flip Flop

Cell culture in 96-well plate for Indigo expression time optimization
Procedure


We set the O.D value to 0.1 and cultured 1.5ml cell in 96 deep well plate. At each time point, we transferred 1ml cell from 96 deep well plate to EP tube and 200λ cell to flat 96 well plate for O.D check.
After cell down in EP tube, we discarded the supernatant and resuspended the pellet in 200λ DMSO. We did sonication to the sample and after another cell down, we took the supernatant for TECAN measurement. Also, we checked absorbance after 3 days incubation and before 3days incubation.
We used TECAN at absorbance 620nm to measure the quantity of indigo production

Results

0818Fig1



We subtracted absorbance value of pure LB from value of samples and we set all samples to have same amount of cell using O.D value
Graph shown below is indigo intensity per cell. After 3 days incubation, indigo seems to be degraded and all the samples seem to have almost same indigo level.

0818Fig1


We excluded 0hr data from the graph to see teh rising tendency clearly.


Discussion


This data shows that the cell successfully produces indigo. We can use this gene as a reporter of our module. However, we only checked the increase of indigo. It may decrease after prolonged culture. We should do additional experiments to get the peak in the graph and optimize the production time of indigo.
Back to the Calendar
October 19th 2012

No Special Event!
Back to the Calendar
October 20th 2012

No Special Event!
Back to the Calendar
October 21st 2012

No Special Event!
Back to the Calendar
October 22nd 2012

No Special Event!
Back to the Calendar
October 23rd 2012

No Special Event!
Back to the Calendar
October 24th 2012

No Special Event!
Back to the Calendar
October 25th 2012

No Special Event!
Back to the Calendar
October 26th 2012

No Special Event!
Back to the Calendar
October 27th 2012

No Special Event!
Back to the Calendar
October 28th 2012

No Special Event!
Back to the Calendar
October 29th 2012

No Special Event!
Back to the Calendar
October 30th 2012

No Special Event!
Back to the Calendar
October 31st 2012

No Special Event!
Back to the Calendar


Kaist Footer