Team:Paris Saclay/Notebook/July/11

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For régulation system:
For régulation system:
-
*1.For those transformation products, RBS+LacS+terminator in PSB1C3 plasmid, the liquid culture has been performed for further experiments. The oligopetides for PCR amplification was made bioinformaticly. The transformation for the terminator BBa_B0010 was still fail.
+
*1.For those transformation products(of yesterday), RBS+LacS+terminator in PSB1C3 plasmid, the liquid culture has been performed for further experiments. The oligopetides for PCR amplification was made bioinformaticly. The transformation for the terminator BBa_B0010 was still fail.
-
*2. The plasmid which contains fnr+RBS+LacZ+terminator and fnr+RBS+AmilCP+terminator was extracted. The restiction digest was performer for them.
+
*2. The plasmid which contains fnr+RBS+LacZ+terminator and fnr+RBS+AmilCP+terminator was extracted. The restiction digest was performeed for them.
For sensor system:
For sensor system:
*3. The selection of bacterian colonies was achieved by analyzing PCR product. The baterian selected are BphR1 c5 and c6; BphR2 c3 and c4; BphA1 c5 and c6,c7,c8.
*3. The selection of bacterian colonies was achieved by analyzing PCR product. The baterian selected are BphR1 c5 and c6; BphR2 c3 and c4; BphA1 c5 and c6,c7,c8.
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=='''Lab work'''==
=='''Lab work'''==
-
Verification by digestion
+
A.aero/anaerobic regulation system:
-
Simple
+
*BioBrick RBS+LacZ+terminator in plasmid PSB1C3
-
DNA 3µl
+
-
Buffer 3 µl
+
-
Enzyme(Xho I or Sac II) 1µl
+
-
H2O 23µl
+
-
Total:30µl
+
-
Double
+
-
DNA 5µl
+
-
Buffer 3 µl
+
-
Enzyme(Xho I or Sac II) 2µl
+
-
H2O 20µl
+
-
Total:30µl
+
-
Buffer :
+
<u>Transformation for BBa_I732019 terminator</u>
-
Ecor I+ PST I -> orange
+
<p>After ong night culture, we observed 2 tiny colonies on the medium. We continued the experiments by performing another liquid culture at 37°C with ampicillin.</p>
-
XhoI->green
+
-
Sac I->blue
+
-
xhoI+Sac II-> green
+
-
size estimed
+
<p>Transformation for BBa_B0010 was always no go</p>
-
Ecor I+PST I : 1069bp and 2750 bp
+
-
Xho I : 2976bp+842bp
+
-
Sac II:3819bp
+
-
Xho I+Sac II: 843bp,616bp,2367bp
+
-
Size observed
+
-
Ecor I+PST I : 2700bp and 1000bp
+
-
Xho I : 4000bp
+
-
Sac II:4000bp
+
-
Xho I+Sac II: 1200bp and 2000bp
+
 +
 +
 +
<u>Verification the transformation of BBa_I004450 in PSB3K3 by digestion and eletrophoresis</u>
 +
 +
<p>We performed 2 types of digestion:</p>
 +
<p align="left">Simple digestion:
 +
*DNA: 3µl
 +
*Buffe:r 3 µl
 +
*Enzyme: 1µl
 +
*H2O: 23µl
 +
*Total: 30µl</p>
 +
 +
<p>Double digestion:
 +
*DNA: 5µl
 +
*Buffer: 3 µl
 +
*Enzyme: 2µl
 +
*H2O: 20µl
 +
*Total: 30µl</p>
 +
 +
<p>Buffer used:
 +
*Ecor I+ PST I -> orange
 +
*Xho I -> green
 +
*Sac II -> blue
 +
*XhoI+Sac II -> green</p>
 +
 +
<p>After the digestion, we performed a eletrophoresis for verification:</p>
 +
 +
{| align="center"
 +
| style="width:350px;border:1px solid black;" | [[File:PSPCR110713d.jpg|center|350px]]
 +
| style="width:350px;border:1px solid black;" |
 +
*Well 1,7 :XhoI+Sac II
 +
*Well 2,8 :Sac II
 +
*Well 3,9 :Ecor I+ PST I
 +
*Well 4,10 :Xho I
 +
*Well 5,11 : control no digested
 +
*gel 0.8%
 +
|}<br>
 +
 +
 +
<p>Estimed size and observed size:</p>
 +
{|border="1" align="center"
 +
|-
 +
|enzyme
 +
|estimed size
 +
|observed size
 +
|-
 +
|Ecor I+PST I
 +
|1069bp and 2750 bp
 +
|1000bp and 2700bp
 +
|-
 +
| Xho I
 +
|2976bp+842bp
 +
|4000bp
 +
|-
 +
|Sac II
 +
|3819bp
 +
|4000bp
 +
|-
 +
|Xho I+Sac II
 +
|843bp,616bp,2367bp
 +
|1200bp and 2000bp
 +
|}

Revision as of 01:13, 22 September 2013

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Notebook : July 11

Summary:

For régulation system:

  • 1.For those transformation products(of yesterday), RBS+LacS+terminator in PSB1C3 plasmid, the liquid culture has been performed for further experiments. The oligopetides for PCR amplification was made bioinformaticly. The transformation for the terminator BBa_B0010 was still fail.
  • 2. The plasmid which contains fnr+RBS+LacZ+terminator and fnr+RBS+AmilCP+terminator was extracted. The restiction digest was performeed for them.

For sensor system:

  • 3. The selection of bacterian colonies was achieved by analyzing PCR product. The baterian selected are BphR1 c5 and c6; BphR2 c3 and c4; BphA1 c5 and c6,c7,c8.

Lab work

A.aero/anaerobic regulation system:

  • BioBrick RBS+LacZ+terminator in plasmid PSB1C3

Transformation for BBa_I732019 terminator

After ong night culture, we observed 2 tiny colonies on the medium. We continued the experiments by performing another liquid culture at 37°C with ampicillin.

Transformation for BBa_B0010 was always no go


Verification the transformation of BBa_I004450 in PSB3K3 by digestion and eletrophoresis

We performed 2 types of digestion:

Simple digestion:

  • DNA: 3µl
  • Buffe:r 3 µl
  • Enzyme: 1µl
  • H2O: 23µl
  • Total: 30µl

Double digestion:

  • DNA: 5µl
  • Buffer: 3 µl
  • Enzyme: 2µl
  • H2O: 20µl
  • Total: 30µl

Buffer used:

  • Ecor I+ PST I -> orange
  • Xho I -> green
  • Sac II -> blue
  • XhoI+Sac II -> green

After the digestion, we performed a eletrophoresis for verification:

PSPCR110713d.jpg
  • Well 1,7 :XhoI+Sac II
  • Well 2,8 :Sac II
  • Well 3,9 :Ecor I+ PST I
  • Well 4,10 :Xho I
  • Well 5,11 : control no digested
  • gel 0.8%


Estimed size and observed size:

enzyme estimed size observed size
Ecor I+PST I 1069bp and 2750 bp 1000bp and 2700bp
Xho I 2976bp+842bp 4000bp
Sac II 3819bp 4000bp
Xho I+Sac II 843bp,616bp,2367bp 1200bp and 2000bp



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