Team:SDU-Denmark/labwork/Notebook/week2

Laboratory Notebook

Here you find the log book for the procedures carried out in the laboratory, starting from week 27.

06-07-2012:

PCR of SST and FFT

We did a polymerase chain reaction with two different settings; one where we used the recommended settings from the protocol and one where we specified the temperature for each cycle in the PCR which should be more optimal for the primers.
The genomic template we used was MCF7. The PCR samples were processed over night (O.N.).

10-07-2012:

Gel for amplification of FFT and SST

We did a gel on the PCR products from O.N. and got some usable bands from the FFT-gene.
We cut the band out of the gel a made a purification of it. We had some problems with the SST-gene and ended with no useable results.
We made a NanoDrop of the purified FFT-gene and got a concentration of -1.0ng/ul. We did a NanoDrop on the PCR products from the FFT-gene before we ran a gel on it to compare with the purified FFT-gene. This gave us a concentration at 103,2ng/ul.
With this result we decided to repeat the purification for the FFT-gene.
We tried to optimise the concentration of the FFT-gene from the gel and ended up with 13,8ng/ul determined by the NanoDrop.

We prepared another PCR for the SST-gene where we adjusted variables like raising the temperature to 63 degrees C for the annealing-step and adjusted the extension time.
PCR was run overnight.