Team:Goettingen/week20-2

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Revision as of 14:22, 16 September 2012

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V09_10_1 Miniprep of pSB1C3 (including motA, motB or yhjH)

Experiment:
Plasmids were prepped using PeqGOLD MiniPrep Kit (Peqlab) accirding to the manual.

V09_10_2 Test digestion of pSB1C3 (including motA, motB or yhjH)

Experiment:
Test digestion of pSB1C3 (including motA, motB or yhjH) with the restriction enzymes EcoRI and PstI. Afterwards, the product was loaded in a 1% analytical gel in order to verify its success.
Observations & Results:
The digestion worked for all clones since fragments with the appropriate size were produced over all.

V09_10_3 Digestion of motA, motB, yhjHand the promoters 20E, 20I and 18C

Experiment:
motA, motB or yhjH were digested with XbaI and PstI. The plasmids with the different promoters were treated with SpeI and PstI. The digestion was then controlled via gel-electrophoresis.
Observations & Results:
Since for all clones band of the expected size could be obtained these were isolated from the gel and purified using PeqGOLD Gelextraction Kit (Peqlab) according to the manual.

V09_10_4 Quickchange of fliC

Experiment:
Quick Change overlapping PCR was performed using designed primers and Pfu Turbo polymerase (see Quick Change protocol in "protocols").
Observations & Results:
Once again the QC PCR failed for no PCR product was observed.

V09_11_1 Ligation and Transformation of motA, motB and yhjH

Experiment:
After the ligation of motA, motB and yhjH into the plasmid J61002 with different promoters (20E, 20I and 18C), the ligation products were transformed into the E. coli strain DH10B.

V09_11_2 Quickchange of fliC part 1

V09_12_1 Quickchange of FliC part 2

V09_12_2 Restriction and Ligation of promoter-RFP constructs into pSB1C3

V09_13_1 Transformation of Psb1c3-Promoter-RFP constructs into e. coli DH10B

V09_13_2 Restriction and Ligation of FliC

V09_13_3 Preparation of over night cultures

Important pages:
Home; Team; Official Team Profile; Project; Parts submitted to the Registry; Modeling; Notebook; Saftey; Attributions

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 <ul>
 <li>Experiment: <br>  Text</li>
+</ul>
+<br>
+<br></td></tr>
+</table>
+<br>
+<table cellpadding="20 px" border="1" bordercolor="black" valign="top">
+<tr bordercolor="black" valign="top">
+<td width="900" bordercolor="black" valign="top">
+<h2><b>V09_13 </b></h2><br>
+<b>V09_13_1 Transformation of Psb1c3-Promoter-RFP constructs into e. coli  DH10B </b><br>
+<ul>
+<li>Experiment: <br>  Text</li>
+</ul>
+<br>
+<b>V09_13_2 Restriction and Ligation of FliC</b><br>
+<ul>
+<li>Experiment: <br>  Text</li>
+</ul>
+<br>
+<b>V09_13_3 Preparation of over night cultures</b><br>
+<ul>
+<li>Experiment: <br> Over night cultures were prepared:<br>
+Psb1c3-MotA<br>
+Psb1c3-MotB<br>
+Bl21<br>
+DH10B<br></li>
 </ul>
 <br>