Team:Goettingen/week6-3

From 2012.igem.org

(Difference between revisions)
 
(One intermediate revision not shown)
Line 29: Line 29:
<b>Preparation of pUC18_TAR_QC</b><br>
<b>Preparation of pUC18_TAR_QC</b><br>
<ul>
<ul>
-
<li>Experiment: <br>1% agarose gel was performed to analyze <i>Dpn</i>I digest. Gel extraction was performed as described before with PeqGOLD Gel extraction kit (Peqlab). Samples were stored at -20 °C.
+
<li>Experiment: <br>1% agarose gel was performed to analyze <i>Dpn</i>I digest. Gel extraction was performed as described before with peqGOLD Gel extraction kit (Peqlab). Samples were stored at -20 °C.
</li>
</li>
</ul>
</ul>
Line 72: Line 72:
<ul>
<ul>
<li>Experiment:  <br>
<li>Experiment:  <br>
-
Minipreps were done with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual. A test digestion was performed using <i>Xba</i>I and the expected fragment of 4342 bp was observed on the corresponding gel. Minipreps were stored at -20 °C. </li>
+
Minipreps were done with peqGOLD Miniprep Kit (Peqlab) according to the user manual. A test digestion was performed using <i>Xba</i>I and the expected fragment of 4342 bp was observed on the corresponding gel. Minipreps were stored at -20 °C. </li>
</ul>
</ul>
<br></td></tr>
<br></td></tr>

Latest revision as of 11:25, 22 September 2012

Deutsch  / English 

#3 Chemoreceptor Library - 6th Week

Back to overview

V06_04


Preparation of pUC18_TAR_QC
  • Experiment:
    1% agarose gel was performed to analyze DpnI digest. Gel extraction was performed as described before with peqGOLD Gel extraction kit (Peqlab). Samples were stored at -20 °C.


V06_05


Transforamtion of DH10B with pUC18_TAR_QC
  • Experiment:
    Transformation was performed according to protocol.


V06_06


Preparation of Miniprep cultures
  • Experiment:
    Preparation of overnight cultures for Miniprep of pUC18_TAR_QC.


V06_07


V06_07_1 Preparation of chemocompetent cells of Δtar
  • Experiment:
    Chemocompetent cells were prepared as described in this protocol. ON cultures from 06.06. were used.

V06_07_2 Miniprep of pUC18_TAR_QC
  • Experiment:
    Minipreps were done with peqGOLD Miniprep Kit (Peqlab) according to the user manual. A test digestion was performed using XbaI and the expected fragment of 4342 bp was observed on the corresponding gel. Minipreps were stored at -20 °C.


V06_08


V06_08_1 Preparative double digest of pUC18_TAR_QC
  • Experiment:
    Digest was performd with XbaI and PstI (according to BioBrick standard) according to protocol. A gelelectrophoresis of the digest was performed which showed the expected bands at 2662 bp and 1680 bp. The insert TAR_QC (1680 bp) was cut from the gel and a gel extraction was performed as described before. Stored at -20 °C.

V06_08_2 Sequencing of pUC18_TAR_QC
  • Experiment:
    Sequencing reactions were done by Prof. Daniels Laboratory (University of Göttingen), Göttingen Genomics Lab and set up according to their protocol. Primers used: pUC18_M13 for, pUC18_M13 rev, pUC18_TAR_Seqfor01, pUC18_TAR_Seqfor02. Dilutions 100 μM, 10 μM, 5 pmol.


Back to overview

↑ Back to top!