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Team:Newcastle/Labwork/3 September 2009
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When we ran the products on a gel. | When we ran the products on a gel. | ||
| - | == Promoter Library Sub-Project == | + | == <u>Promoter Library Sub-Project</u> == |
=== Introduction === | === Introduction === | ||
From the previous lab session the products of the PCR reactions attempting to amplify the ''sigA'' promoter library (synthesized sequences of the ''sigA'' library containing degeneracies) were run through 1.5% agarose gel in electrophoresis. Analysis of the gel showed that although there were hazy bands forming, these may have not been the result of the PCR products and may have been caused by the primer-dimer complexes formed. The promoter fragments are roughly 100bp however, meaning that there may be little distinction between the promoters and the primer-dimers in 1.5% agarose gel! In light of this experiment, '''today's task will involve cleaning up the PCR products for another run of DNA gel electrophoresis tomorrow'''. | From the previous lab session the products of the PCR reactions attempting to amplify the ''sigA'' promoter library (synthesized sequences of the ''sigA'' library containing degeneracies) were run through 1.5% agarose gel in electrophoresis. Analysis of the gel showed that although there were hazy bands forming, these may have not been the result of the PCR products and may have been caused by the primer-dimer complexes formed. The promoter fragments are roughly 100bp however, meaning that there may be little distinction between the promoters and the primer-dimers in 1.5% agarose gel! In light of this experiment, '''today's task will involve cleaning up the PCR products for another run of DNA gel electrophoresis tomorrow'''. | ||
Revision as of 23:37, 5 September 2009
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Lab Work - 03/09/09
Chassis team
Today we continued the lab work on our PCR primers. We again attempted to use two pairs of our primers; the ones to amplify the sleB and cwlJ genes from the B. subtilis genome. We prepared the solutions needed, and followed the PCR protocol. We rehydrated our primers, and diluted them accordingly. The TMs were lowered, following yesterday's failed attempt to 50 and 45 degrees. We prepared four lots of solutions, two with genomic DNA and two without (our controls), containing 2 and 3ul of primer. They were loaded into the PCR machine, which ran for around 2 hours 23 minutes.
When we ran the products on a gel.
Promoter Library Sub-Project
Introduction
From the previous lab session the products of the PCR reactions attempting to amplify the sigA promoter library (synthesized sequences of the sigA library containing degeneracies) were run through 1.5% agarose gel in electrophoresis. Analysis of the gel showed that although there were hazy bands forming, these may have not been the result of the PCR products and may have been caused by the primer-dimer complexes formed. The promoter fragments are roughly 100bp however, meaning that there may be little distinction between the promoters and the primer-dimers in 1.5% agarose gel! In light of this experiment, today's task will involve cleaning up the PCR products for another run of DNA gel electrophoresis tomorrow.
PCR Product Cleanup
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)
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