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Team:Paris Saclay/Notebook/August/19
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==== Digestion of PCR results ==== | ==== Digestion of PCR results ==== | ||
Revision as of 09:06, 14 October 2014
Contents |
Tuesday 19th August
Lab work
Chromoprotein + Lemon scent
Electrophoresis with the PpsI plasmide (obtain on thursday) + the chromoprotein PCR: we obtain results as expected
- ladder 10µl
- pPSI 1.2µl
- pPSI 1.2µl
- pPSI 1.2µl
- pPSI 1.2µl
- pPSI 1.2µl
- p cola 1.2µl
- PCR chromoprotein 2µl
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 |
| Antibiotic | Concentration |
|---|---|
| Ampicillin | 4*10-4 |
Bacterial culture
by Laetitia
After the bacterial transformation, we spread for each strain (LS or PS or GS):
| dish type | 100µl | 200µl | concentrate | control |
|---|---|---|---|---|
| Ampicillin | X | |||
| Ampicillin+IPTG+XGal | X | X | X | X |
We leave the dishes overnight at 37°C.
