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Team:UCL/Labbook
From 2013.igem.org
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<table> | <table> | ||
<tr> | <tr> | ||
| - | <th> | + | <th>Vial</th> |
| - | <th> | + | <th>Ampicillin Plate</th> |
| + | <th>Plasmid Insertion</th> | ||
| + | <th>Colony Count</th> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td> | + | <td>1 (Main)</td> |
| - | <td> | + | <td>Yes</td> |
| + | <td>Yes</td> | ||
| + | <td>0</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td> | + | <tr> |
| - | <td> | + | <td>2 (Positive Control)</td> |
| + | <td>No</td> | ||
| + | <td>Yes</td> | ||
| + | <td>100+</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>3 (Negative Control)</td> | ||
| + | <td>Yes</td> | ||
| + | <td>No</td> | ||
| + | <td>0</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
Revision as of 11:07, 9 September 2013
June
Week 1-3
No lab work
Week 4
26th June - We are given safety training in the Advanced Centre for Biochemical Engineering in all relevant laboratories, as well as general procedures in case of emergency.
July
Week 5-6
No lab work
Week 7
Bacterial Lab
15th July - The team is introduced to the laboratories which will be used during the summer for both bacterial and mammalian experiments. 16th July - 5X M9 salts [link to protocol], minimal agar [link to protocol], 1.4% molten agar solution [link to protocol] and 0.1M CaCl2/15% glycerol [link to protocol] were prepared for the generation of competent cells. Minimal agar plates were poured and streaked [link to streaking protocol] with W3110 Escherichia coli cells and left overnight to incubate at 37C. 17th July - Very little colony growth was observed from W3110 E.coli streaked plates. Plates were therefore left to incubate for a further 17 hours. 18th July - Sufficient colony growth allowed for the selection of a single colony from each plate. This was then inoculated in 5ul LB media [link to LB recipe] + 100ul 1M MgSO4 and left to incu-shake overnight at 37C. 19th July - Cultures re-suspended in new LB media and 100µl aliquots placed into individual eppendorf tubes for placement at -80C. Mammalian Lab 17th July - Mammalian cell culture and maintenance [link to mammalian protocol] training by Mrs. Ludmilla Ruban. Passaged primary MEF (mouse embryonic fibroblast) cells. Passage 3. 18th July - MEF passage 4 19th July - MEF passage 5Week 8
Bacterial Lab
22nd July -Transformation [link to transformation protocol] of our competent cells with plasmid YB3110 was carried out. 23rd July - No colony growth was observed on Ampicillin plates [link to ampicillin plate protocol] indicating no plasmid uptake. Transformation was repeated with YB3110.| Vial | Ampicillin Plate | Plasmid Insertion | Colony Count |
|---|---|---|---|
| 1 (Main) | Yes | Yes | 0 |
| 2 (Positive Control) | No | Yes | 100+ |
| 3 (Negative Control) | Yes | No | 0 |
Week 9
Bacterial Lab
August
