Team:Paris Saclay/Notebook/September/2

Labwork

B-Contruction of the fusion protein

By Hv

We can see that the blue colonies of E.coli are not blue in a simple petri dish with ampicillin. It's a fail. So, to figure our problem we decided to do a sequencing of our plasmids.

We chose 6 colonies from an Xgal-IPTG petri dish:

- 1 completely blue

- 2 blue with a white ring around the blue

- 3 white colony

PS: we found that there is 2 types of blue colonies: completely blue and blue with a white ring around the blue)

We first did a liquid culture in 5 mL of LB.

D- Lemon scent

By Melanie

Limonene synthase

Migration of the PCR done yesterday

picture 1 well 1= ladder well 2-6 = pPS5 other well = LS PCR

picture 2 = LS PCR As we see, there are nothing on our gel so we conclude that we failed to clone LS in pGMETeasy

pPPS5

the photo in the same as previously: We saw a band but we will check another time by PCR to be sure.

pPS3 and pPS4

Plasmid extraction using the plasmid DNA extraction kit and electrophoresis:

We conclude that we have done a good manipulation.

Limonene synthase PCR

As we saw that we don't have anything for LS, We try to do other PCR from the Biobrick (BBA762100). We use two enzyme: Dream taq and Vent taq

Tubes were placed in PCR machine with the following parameters.

pPS5

To have more plasmid We do some bacteria culture (pPS5)