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Team:Paris Saclay/Notebook/August/19
From 2014.igem.org
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 |
|---|---|
| PCR 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 dish
by Terry
10 petri dish using :
- LB+Agar
-IPTG(1M) at 0,5mM
-80µg of X-Gal(80mg/mL)
- Amp (1/1000)
4 petri dish using :
- LB+Agar
- Amp (1/1000)
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
