Team:KULeuven/29 July 2008

Wet Lab


Only one ligation succeeded (R0084+B0032). We think that the others failed because the vectors weren't properly cut. Therefore, we did another digest. This time we cut the vector with Xba I, and only when this worked, we cut it with EcoR I. We did this because Xba I has problems with cleaving close to the ends of DNA fragments. The problem is that Xba I doesn't seem to cut at all. Perhaps there's something wrong with the enzyme. We are trying to overcome this problem by allowing the restriction with Xba I to continue overnight.

Some more plasmids were isolated from the cells using the Qiagen Miniprep Kit. It concerns parts used in the filter and in the inverter (J23022, J23109, J23032, I712074, B0015, R0084, B0032 and B0034, C0012, R0011, F1610). We also digested some of these isolated plasmids. The ones that were cut with EcoR I and Spe I succeeded (J23109, I712074 and R0084) and were cut out of the agarose gel and were purified. The ones that were cut with Xba I failed (J23022, J23032 and B0015) - we didn't get the expected result with gel electrophoresis.

General
GFP primers were finished and ordered. Antisense LuxI primers were written down and will soon be ordered. The primers have been given their place in the notebook section of the wiki.

As we noticed problems with M30109 - the "physical DNA" description of the part on the Registry proves nothing good and Melbourne 2007 reported problems with the part - we contacted the Melbourne 2008 team. They work with this part, and we hope that they can help us out.

We did some research about the origin of the EnvZ::KmR mutation in V1012 and found the paper of Mizuno and Mizushima. Based on the information in that paper, we will make primers to check whether the transduction has been a succes (or not).

Modeling
Mainly chillin' out, memory was finished and complete model is functional.

Wiki
Modeling section got taken care of some more, work on new dropdown menu proceeds...

Quote Of The Day
Sigrid: Het is te hopen dat de getransduceerde cellen een groeischeut krijgen. Stefanie: Daar wacht ik nu al 22 jaar op!