Team:Goettingen/week10-2

From 2012.igem.org

(Difference between revisions)

Revision as of 14:52, 16 September 2012

Language:

English,

Deutsch

#2 Speed Improvement - 10th week

Back to overview

V07_02

V07_02_1 Preparative double digestion of 20E-flhDC, 2G-flhDC, 18O-flhDC and pSB1C3

Experiment:
In order to clone the flhDC-promoter constructs intro pSB1C3, all components were digested with EcoRI and PstI according to the protocol. The restrictio products were subsequently separated on a gel in order to verify the digestions success and purification of the desired fragments.

Observations & Results:
The gel did not feature fragments of the expected size.Obviously, something was wrong with the digestion.

V07_02_2 Preparation of over night cultures

Experiment:
In order to gain more plasmid material of the flhDC-promoter constructs over night cultured were prepared for a subsequent plasmid isolation.
20E - #2
20G - #1
2G - #1
20I - #2
18M - #2
18O - #2
18K - #1
18C - #2

V07_03

V07_03_1 Miniprep of the flhDC-promoter constructs

Experiment:
Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.

V07_03_2 Amplification of the genes fliC (DH10B), fliC (Salmonella), motA, motB and yhjH

Experiment:
The genes fliC (DH10B), fliC (Salmonella), motA, motB and yhjH were amplified by Pfu-Polymerase according to the following protocol. The success of the PCR was subsequently investigated preparing a 1% agarose gel.
Observations & Results:
The PCR failed for no bands were visible on the gel whereas the marker was clearly discernible.

V07_04

Repetition of the amplification of fliC (DH10B), fliC (Salmonella), motA, motB and yhjH

Experiment:
The genes fliC (DH10B), fliC (Salmonella), motA, motB and yhjH were amplified by Pfu-Polymerase according to the following protocol. Additionally, a negative control as well as a positive control were prepared this time. The success of the PCR was subsequently investigated preparing a 1% agarose gel.
Observations & Results:
The did not function again for no bands were visible on the gel; only the marker was clearly discernible. However, since also the positive control did not feature the slightest band, we assume that something might be wrong with our polymerase.

V06_15

Miniprep of pBAD-sfGFP

Experiment:
Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.

↑ Return to top

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

@@ Line 565: / Line 565: @@
 Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.</li></ul>
 <br>
-<b>V07_03_2 Amplification of the genes <i>fliC</i> (DH10B), <i>fliC</i> (Salmonella), <i>motA</i>, <i>motB</i> and <i>yhjH</i></b><br>
+<b>V07_03_2 Amplification of the genes <i>fliC</i> (DH10B), <i>fliC</i> (<i>Salmonella</i>), <i>motA</i>, <i>motB</i> and <i>yhjH</i></b><br>
 <ul>
 <li>Experiment:  <br>
-The genes <i>fliC</i> (DH10B), <i>fliC</i> (Salmonella), <i>motA</i>, <i>motB</i> and <i>yhjH</i> were amplified by Pfu-Polymerase according to the following protocol. The success of the PCR was subsequently investigated preparing a 1% agarose gel.<br>
+The genes <i>fliC</i> (DH10B), <i>fliC</i> (<i>Salmonella</i>), <i>motA</i>, <i>motB</i> and <i>yhjH</i> were amplified by Pfu-Polymerase according to the following protocol. The success of the PCR was subsequently investigated preparing a 1% agarose gel.<br>
 <li>Observations & Results: <br>
-The PCR failed for no bands were visible on the gels whereas the marker was clearly discernible.</li>
+The PCR failed for no bands were visible on the gel whereas the marker was clearly discernible.</li>
 </ul><br>
@@ Line 581: / Line 581: @@
 <tr bordercolor="black" valign="top">
 <td width="900" bordercolor="black" valign="top">
-<h2><b>V06_14 </b></h2><br>
+<h2><b>V07_04 </b></h2><br>
-<b> Preparation of over night cultures</i></i></b><br>
+<b> Repetition of the amplification of <i>fliC</i> (DH10B), <i>fliC</i> (<i>Salmonella</i>), <i>motA</i>, <i>motB</i> and <i>yhjH</i> </i></i></b><br>
 <ul>
 <li>Experiment: <br>
-Over night cultures of the pBAD-<i>sfGFP</i> clones were prepared in order to isolate the plasmids. <br>
+The genes <i>fliC</i> (DH10B), <i>fliC</i> (<i>Salmonella</i>), <i>motA</i>, <i>motB</i> and <i>yhjH</i> were amplified by Pfu-Polymerase according to the following protocol. Additionally, a negative control as well as a positive control were prepared this time. The success of the PCR was subsequently investigated preparing a 1% agarose gel.<br></li>
+<li>Observations & Results: <br>
+The did not function again for no bands were visible on the gel; only the marker was clearly discernible. However, since also the positive control did not feature the slightest band, we assume that something might be wrong with our polymerase.</li>
 <br></td></tr>
 </table>