Team:Paris Saclay/Notebook/August/19

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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 :

- 250mL LB+Agar

-250 µL X-Gal(80mg/mL)

- 250 µL ampi

- 125 µL IPTG

-->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

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