Team:Newcastle/13 August 2010

From 2010.igem.org

Revision as of 01:02, 26 October 2010 by Alankoh (Talk | contribs)

iGEM Homepage Newcastle University BacillaFilla Homepage Image Map



Contents

Gel electrophoresis of subtilin immunity BioBrick fragments

Aims

The aim of this experiment is to do gel extraction for the spaIFEG gene cluster amplifed on the 12th August, 2010 and the Plasmid Vector, Promoter & RBS and Double terminator amplifed on the 11th August, 2010. Finally, NanoDrop will be performed for all the fragments.

Materials and protocol

Please refer to the gel extraction and NanoDrop protocols. Instead of using the centrifuge to bind the DNA from our sample to the QIAquick column, a vacuum manifold was used instead. It is the first time it had been used. The advantage of this was that the extraction could be carried out all at the same time.

Gel Extraction of spaIFEG genes Setting up the tubes for the vacuum manifold Setting up the tubes for the vacuum manifold Vacuum manifold with the QIAquick spin columns loaded with the samples

Results

The results from the Nanodrop Spectrophotometer of the four parts are as follows:

  • Vector - 21.3 ng/μl
  • Promoter - 28.8 ng/μl
  • Coding sequence - 27.0 ng/μl
  • Terminator - 27.5 ng/μl

Discussion

We chose the three spaIFEG gene (part 3) PCR tubes, using Tms 51°C, 56°C and 61°C because they were the mid-ranged Tms. Gel extraction was then performed, by firstly cutting the gels under U.V light, and then apply the protocol for completion.

Conclusion

The results from the NanoDrop reading suggests that our results were positive, which indicates that we have successfully extracted all the parts required for the Subtilin Immunity BioBrick. Gibson cloning method will be used next week.



Go back to our main Lab book page


Newcastle University logo.png    Newcastle cbcb logo.pngNewcastle Biomedicine logo.gif    Team Newcastle CEG logo.gif
Newcastle iww logo.jpg  UNIPV Pavia Logo.gif  Newcastle BBSRC.gif    Newcastle Genevision logo.png Newcastle WelcomeTrust.jpg
FaceBook Icon