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Team:Paris Saclay/Notebook/August/19
From 2014.igem.org
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We transformed 100µL of competent bacteria and we added the totality of the ligation product (pGEMTeasy containing LS or PS or GS) | We transformed 100µL of competent bacteria and we added the totality of the ligation product (pGEMTeasy containing LS or PS or GS) | ||
| - | ====Preparation of petri | + | ====Preparation of petri dishes==== |
''by Terry'' | ''by Terry'' | ||
| - | --> | + | {| class="wikitable centre" width="50%" |
| + | |+ 10 dishes with: | ||
| + | |- | ||
| + | ! scope=col | Antibiotic | ||
| + | ! scope=col | Concentration | ||
| + | |- | ||
| + | |XGal | ||
| + | |80μg/ml | ||
| + | |- | ||
| + | |IPTG | ||
| + | |2*10<sup>-3</sup> | ||
| + | |- | ||
| + | |Ampicillin | ||
| + | |10<sup>-3</sup> | ||
| + | |}--> | ||
| - | + | '''4 petri dish using:''' | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
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- 100mL LB+Agar | - 100mL LB+Agar | ||
Revision as of 09:06, 22 August 2014
Contents |
Tuesday 19th August
Lab work
B and D
Electrophoresis with the PpsI plasmide (obtain on thursday) + the chromoprotein PCR: we obtain results as expected
D - Lemon Scent
Digestion of PCR results
by Sean, Hoang Vu, Eugene
| components | volumes |
|---|---|
| PCR purified LS | 15 μl |
| FastDigest green buffer 10X | 2 μl |
| PacI | 1 µl |
| H2O | 2 µl |
| components | volumes |
|---|---|
| PCR purified GS | 15 μl |
| FastDigest green buffer 10X | 2 μl |
| PacI | 1 µl |
| H2O | 2 µl |
| components | volumes |
|---|---|
| PCR purified PS | 15 μl |
| FastDigest green buffer 10X | 2 μl |
| PacI | 1 µl |
| H2O | 2 µl |
| components | volumes |
|---|---|
| Purified pPS1 | 30 μl |
| FastDigest green buffer 10X | 4 μl |
| PacI | 2 µl |
| H2O | 4 µl |
Purification of PCR products of limonen synthase
by Laetitia
We used the PCR products of limonen synthase prepared previously : August 18th
The five samples were pooled in one and then purified using this protocol : protocol
Bacterial transformation
by Laetitia
We used DH5alpha competent bacteria (prepared the 14/08) and they were transformed using the ligation product prepared previously August 18th using this protocol : protocol
We transformed 100µL of competent bacteria and we added the totality of the ligation product (pGEMTeasy containing LS or PS or GS)
Preparation of petri dishes
by Terry
| Antibiotic | Concentration |
|---|---|
| XGal | 80μg/ml |
| IPTG | 2*10-3 |
| Ampicillin | 10-3 |
4 petri dish using:
- 100mL LB+Agar
- 100 µL ampi
Bacterial culture
by Laetitia
After the bacterial transformation, we spread for each strain (LS or PS or GS) :
-100µL on LB-Agar+Amp
-100µL on LB-Agar+Amp+IPTG+X-Gal
-200µL on LB-Agar+Amp+IPTG+X-Gal
-100µL of concentrated bacteria on LB-Agar+Amp+IPTG+X-Gal
37°C at night
