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Team:Paris Liliane Bettencourt/Notebook/2011/09/07/
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* YFP TetR BB -> SpeI site are not there in the 2 clones sequenced | * YFP TetR BB -> SpeI site are not there in the 2 clones sequenced | ||
* ComS -> Ok on the two clones sequenced | * ComS -> Ok on the two clones sequenced | ||
| + | == Hovannes-Baptiste == | ||
| + | === Preparation of slides === | ||
| + | |||
| + | Dilution of overnight cultures : PY79 (gfp-) and PY79 (gfp+) . <br> | ||
| + | |||
| + | |||
| + | We waited to an OD of 0.4 (600 nm). | ||
| + | |||
| + | Two well slides : 1-control (PY79 only) 2-Mix (both strains) | ||
| + | <br> | ||
| + | |||
| + | === Observation === | ||
| + | -37°C Microscopy-<br> | ||
| + | |||
| + | We observed the plate with TRANS and YFP-filter settings on the Nikon microscope. We quickly saw that our PY79 + S12 gene was not correct since we found only one cell producing GFP. We will retry the experiment tomorrow with the other -20°C glycerol for this strain, hoping it is not contaminated. | ||
| + | |||
| + | [[File:0907_trans_PY79_PY79S12.jpg|500px|thumb|center|TRANS at t=0min]] | ||
| + | [[File:0907_fluo_PY79_PY79S12.jpg|500px|thumb|center|GFP at t=0min]] | ||
| + | |||
| + | |||
| + | We followed the plate a few hours nonetheless but no result was visible (the fluorescent cell divided only 2 times). | ||
Latest revision as of 20:18, 17 September 2011
Contents |
Cyrille
Miniprep and sequencing
Miniprep of:
- pVeg-YFP TetR in pHM3 -> Sequence ok
- TetO pVeg-YFP TetR in pHM3 -> Sequence shows that only TetO was there
- YFP TetR BB -> SpeI site are not there in the 2 clones sequenced
- ComS -> Ok on the two clones sequenced
Hovannes-Baptiste
Preparation of slides
Dilution of overnight cultures : PY79 (gfp-) and PY79 (gfp+) .
We waited to an OD of 0.4 (600 nm).
Two well slides : 1-control (PY79 only) 2-Mix (both strains)
Observation
-37°C Microscopy-
We observed the plate with TRANS and YFP-filter settings on the Nikon microscope. We quickly saw that our PY79 + S12 gene was not correct since we found only one cell producing GFP. We will retry the experiment tomorrow with the other -20°C glycerol for this strain, hoping it is not contaminated.
We followed the plate a few hours nonetheless but no result was visible (the fluorescent cell divided only 2 times).
