Team:Grenoble/Biology/Notebook/July

From 2012.igem.org

(Difference between revisions)
Line 14: Line 14:
<h2> Goal of the week </h2>
<h2> Goal of the week </h2>
We wanted to test the PCR conditions, recover and amplify the biobricks involved in our genetic networks :
We wanted to test the PCR conditions, recover and amplify the biobricks involved in our genetic networks :
-
<ol>
+
<ul>
<li>pAra/Bad_RBS_GFP (1300bp)</li>
<li>pAra/Bad_RBS_GFP (1300bp)</li>
<li>RBS_Cya (2600bp)</li>
<li>RBS_Cya (2600bp)</li>
Line 26: Line 26:
<li>pSB4K5 (2400bp)</li>
<li>pSB4K5 (2400bp)</li>
<li>pSB3C5 (2400bp)</li>
<li>pSB3C5 (2400bp)</li>
-
</ol>
+
</ul>
<h2> Tuesday, July 03<span class="exposant">rd</span>:</h2>
<h2> Tuesday, July 03<span class="exposant">rd</span>:</h2>
Precultured cells are prepared:
Precultured cells are prepared:
-
<ol>
+
<ul>
<li>Strains = BW25113 WT, BW25113 cya- and BW25113 cya- pAra/Bad </li>
<li>Strains = BW25113 WT, BW25113 cya- and BW25113 cya- pAra/Bad </li>
<li>Conditions = LB liquid medium, 37°C, 200rpm, overnight </li>
<li>Conditions = LB liquid medium, 37°C, 200rpm, overnight </li>
-
</ol>
+
</ul>
<h2>Wednesday, July 04<span class="exposant">th</span>:</h2>
<h2>Wednesday, July 04<span class="exposant">th</span>:</h2>
We did PCRs on colony with a GoTaq enzyme (protocol) to amplify:
We did PCRs on colony with a GoTaq enzyme (protocol) to amplify:
-
<ol>
+
<ul>
<li>fha1, RsmA, rsmY and pSB1A3 from iGEM Grenoble 2011 glycerol stock.</li>
<li>fha1, RsmA, rsmY and pSB1A3 from iGEM Grenoble 2011 glycerol stock.</li>
<li>pAra/Bad_RBS_GFP from BW25113 cya- pAra/Bad precultured cells.</li>
<li>pAra/Bad_RBS_GFP from BW25113 cya- pAra/Bad precultured cells.</li>
<li>RBS_Cya from BW25113 WT precultured cells.</li>
<li>RBS_Cya from BW25113 WT precultured cells.</li>
-
</ol>
+
</ul>
<br/>
<br/>
Using iGEM 2012 biobricks we transformed BW25113 WT cells (protocol). We obtained four transformed strains with four different biobricks:
Using iGEM 2012 biobricks we transformed BW25113 WT cells (protocol). We obtained four transformed strains with four different biobricks:
-
<ol>
+
<ul>
<li>BBa_I13601: pLAC_RBS</li>
<li>BBa_I13601: pLAC_RBS</li>
<li>BBa_E0422: eCFP</li>
<li>BBa_E0422: eCFP</li>
<li>pSB3C5 plasmid</li>
<li>pSB3C5 plasmid</li>
<li>psB4K5 plasmid</li>
<li>psB4K5 plasmid</li>
-
</ol>
+
</ul>
<h2>Thursday, July 5<span class="exposant">th</span>:</h2>
<h2>Thursday, July 5<span class="exposant">th</span>:</h2>
Line 54: Line 54:
<br/>
<br/>
In order to separate the PCR products (12/07/04), we prepared two gels:
In order to separate the PCR products (12/07/04), we prepared two gels:
-
<ol>
+
<ul>
<li>a 1.3% Tris base Acetic Acid EDTA (TAE) agarose gel, for the small fragments: RsmA, fha1 and rsmY (length < 1000bp)</li>
<li>a 1.3% Tris base Acetic Acid EDTA (TAE) agarose gel, for the small fragments: RsmA, fha1 and rsmY (length < 1000bp)</li>
<li>a 0.8% TAE agarose gel, for the big fragments: pSB1A3, RBS_Cya, and pAra/Bad_RBS_GFP (length > 1000bp)</li>
<li>a 0.8% TAE agarose gel, for the big fragments: pSB1A3, RBS_Cya, and pAra/Bad_RBS_GFP (length > 1000bp)</li>
 +
</ul>
<br/>Migration conditions = 100V during 35 min.
<br/>Migration conditions = 100V during 35 min.
In order to reveal the fragments, we used Ethidium Bromide (EtBr).
In order to reveal the fragments, we used Ethidium Bromide (EtBr).

Revision as of 13:23, 6 August 2012

iGEM Grenoble 2012

iGEM Grenoble iGEM
iGEM 2012
main page
Project
week 27 week 28 week 29 week 30

Week 27: July 02nd to July 08st

Goal of the week

We wanted to test the PCR conditions, recover and amplify the biobricks involved in our genetic networks :
  • pAra/Bad_RBS_GFP (1300bp)
  • RBS_Cya (2600bp)
  • pLAC (100bp)
  • fha (80bp)
  • eCFP (800bp)
  • pLAC_RBS (120bp)
  • RsmA (200bp)
  • rsmY (170bp)
  • pSB1A3 (2400bp)
  • pSB4K5 (2400bp)
  • pSB3C5 (2400bp)

Tuesday, July 03rd:

Precultured cells are prepared:
  • Strains = BW25113 WT, BW25113 cya- and BW25113 cya- pAra/Bad
  • Conditions = LB liquid medium, 37°C, 200rpm, overnight

Wednesday, July 04th:

We did PCRs on colony with a GoTaq enzyme (protocol) to amplify:
  • fha1, RsmA, rsmY and pSB1A3 from iGEM Grenoble 2011 glycerol stock.
  • pAra/Bad_RBS_GFP from BW25113 cya- pAra/Bad precultured cells.
  • RBS_Cya from BW25113 WT precultured cells.

Using iGEM 2012 biobricks we transformed BW25113 WT cells (protocol). We obtained four transformed strains with four different biobricks:
  • BBa_I13601: pLAC_RBS
  • BBa_E0422: eCFP
  • pSB3C5 plasmid
  • psB4K5 plasmid

Thursday, July 5th:

The transformations (12/07/04) show no growth on LB Agar plate + antibiotics.

In order to separate the PCR products (12/07/04), we prepared two gels:
  • a 1.3% Tris base Acetic Acid EDTA (TAE) agarose gel, for the small fragments: RsmA, fha1 and rsmY (length < 1000bp)
  • a 0.8% TAE agarose gel, for the big fragments: pSB1A3, RBS_Cya, and pAra/Bad_RBS_GFP (length > 1000bp)

Migration conditions = 100V during 35 min. In order to reveal the fragments, we used Ethidium Bromide (EtBr).

Retrieved from "http://2012.igem.org/Team:Grenoble/Biology/Notebook/July"