Team:Newcastle/Labwork/4 September 2009
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Revision as of 01:12, 6 September 2009
Contents |
Lab Work 04/09/09
Promoter Library Sub-Project
Introduction
In yesterday's lab session the products from the PCR reactions on the C0, V1, V2 and V3 samples were purified. The gel electrophoresis carried out on the unpurified samples the previous day showed hazy bands however it was uncertain that these bands were the promoters or just primer-dimers. Today, we aim to carry out DNA gel electrophoresis on these purified samples (in 1.5% agarose gel) to determine whether any PCR amplification reactions worked.
Preparing samples and DNA Gel Electrophoresis
The purified PCR products were treated in a similar way to which DNA samples are treated in the gel electrophoresis protocol except 5ul of each sample was extracted and to each of these 1ul of loading buffer (dye) added. 15ul of HindIII' ladder will be added in the first and last wells of the 1.5% agarose gel. This is how the wells will be organised:
- Lane 1 = blank
- Lane 2 = 15ul of HindIII DNA ladder
- Lane 3 = 6ul of C0 (Sample 1)
- Lane 4 = 6ul of V1 (Sample 2)
- Lane 5 = 6ul of V2 (Sample 3)
- Lane 6 = 6ul of V3 (Sample 4)
- Lane 7 = 15ul of HindIII DNA ladder
The 1.5% agarose was prepared and poured in the same way as the DNA gel electrophoresis protocol suggests. The samples were then added to the wells in the way described above; the voltage was set to 60mv and left for 15-20 minutes. The photograph obtained from the resulting gel can be seen in the 'Results' section.
Results
News
Events
- 20 – 21 June 2009 - Europe workshop (London)
- 23 – 24 June 2009 - UK iGEM meetup (Edinburgh)
- 23 October Practice Presentation (Newcastle)
- 23 October T-shirts are ready
- 27 October Practice Presentation (Sunderland)
- 27 October Poster is ready
- 30 October – 2 November 2009 - Jamboree (Boston)
Social Net
- Newcastle iGEM Twitter
- [http://www.facebook.com/home.php#/group.php?gid=131709337641 Newcastle on Facebook]
- [http://www.youtube.com/user/newcastle2009igem Newcastle Youtube Channel]