Team:Goettingen/week4-2

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#2 Speed Improvement - 4th week

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V05_23

V05_23_1 Preparing BioBrick plasmids with constitutive promoters

Experiment:
J23100 – 18C – x2547
J23114 – 20 I – x256
J23112 – 20 E – x1
resuspended in 10 µl water and stored at -20°C

V05_23_2 Chemical transformation of P1 18C and P1 20I in E. coli (DH10B)

Experiment:
For the chemical transformations the standard protocol was followed.
Observations & Results:
On all plates bacterial growth could be observed, even on the negative control plate. We suggest that the plates are all right, but the competent cells are not. The over night culture seems to have been prepared with cells already containing puc18 and thus an ampicillin resistance. The experiment has to be repeated we new competent cells.

V05_24

V05_24_1 Preparing competent cells of E. coli (DH10B)

Experiment:
The preparation of the competent cells was performed according to the following protocol. This time cells without any resistance were utilized for inoculation

V05_23_2 Chemical transformation (DH10B)

Experiment:
For the chemical transformations the standard protocol was followed. The following constructs were transformed into
E. coli
J23112 - 20E - x1
J23113 - 20G - x21
J23109 - 2G - x106
J23114 - 20I - x256
J23105 - 18M - x623
J23106 - 18O - x1185
J23104 - 18K - x 1831
J23100 - 18 C - x 2547
Observations & Results:
On all plates bacterial growth could be observed, even on the negative control plate. We suggest that the plates are all right, but the competent cells are not. The over night culture seems to have been prepared with cells already containing puc18 and thus an ampicillin resistance. The experiment has to be repeated we new competent cells.

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@@ Line 553: / Line 553: @@
 <tr bordercolor="black" valign="top">
 <td width="900" bordercolor="black" valign="top">
-<h2><b>V05_23 </b></h2><br>
+<h2><b>V05_24 </b></h2><br>
-<b>V05_23_1 Preparing BioBrick plasmids with constitutive promoters</b><br>
+<b>V05_24_1 Preparing competent cells of <i>E. coli</i> (DH10B)</b><br>
 <ul>
 <li>Experiment:  <br>
-J23100 – 18C – x2547 <br>
+The preparation of the competent cells was performed according to the following protocol. This time cells without any resistance were utilized for inoculation</li>
-J23114 – 20 I – x256 <br>
-J23112 – 20 E – x1 <br>
-resuspended in 10 µl water and stored at -20°C </li>
 </ul>
 <br>
-<b>V05_23_2 Chemical transformation of P1 18C and P1 20I in <i>E. coli</i> (DH10B) </b><br>
+<b>V05_23_2 Chemical transformation (DH10B) </b><br>
 <ul>
 <li>Experiment:  <br>
-For the chemical transformations the standard protocol was followed. </li>
+For the chemical transformations the standard protocol was followed. The following constructs were transformed into <br>
+ <i>E. coli</i> <br>
+J23112 - 20E - x1 <br>
+J23113 - 20G - x21 <br>
+J23109  - 2G - x106 <br>
+J23114 - 20I - x256 <br>
+J23105 - 18M - x623 <br>
+J23106 - 18O - x1185 <br>
+J23104 - 18K - x 1831 <br>
+J23100 - 18 C - x 2547 <br> </li>
 <li>Observations & Results: <br>
 On all plates bacterial growth could be observed, even on the negative control plate. We suggest that the plates are all right, but the competent cells are not. The over night culture seems to have been prepared with cells already containing puc18 and thus an ampicillin resistance. The experiment has to be repeated we new competent cells.