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Minnesota/8 July 2008
From 2008.igem.org
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|2. '''Restart/Redo PCR''': Problem with original PCR was an error in the calculation of DNA concentration of product. 07-07-2008 we added only 1.0uL of PCR to digest reactions meaning only 3ng of DNA was in the solution. That was not a substantial amount of DNA to be seen in a electrophoretic gel. FIX - recalculated DNA concentration and is 3ng/1uL instead of 300ng/uL. Redid PCR2 with 100.0ng DNA instead of 1ng of DNA by adding 10.0uL of DNA. Will run gel with new PCR undigested (original PCR was digested). | |2. '''Restart/Redo PCR''': Problem with original PCR was an error in the calculation of DNA concentration of product. 07-07-2008 we added only 1.0uL of PCR to digest reactions meaning only 3ng of DNA was in the solution. That was not a substantial amount of DNA to be seen in a electrophoretic gel. FIX - recalculated DNA concentration and is 3ng/1uL instead of 300ng/uL. Redid PCR2 with 100.0ng DNA instead of 1ng of DNA by adding 10.0uL of DNA. Will run gel with new PCR undigested (original PCR was digested). | ||
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| - | |3. '''PCR''': New PCR completed successfully. The RBS has been incorporated into the lambda cI gene, and the resulting DNA has a concentration of 23 ng/ul. Analysis by agarose gel electrophoresis confirmed that the DNA fragment is of the appropriate size. | + | |3. '''PCR''': New PCR completed successfully. The RBS has been incorporated into the lambda cI gene, and the resulting DNA has a concentration of 23 ng/ul. Analysis by agarose gel electrophoresis confirmed that the DNA fragment is of the appropriate size (~780 bp). Top lane is 1kb ladder, third lane is PCR product. |
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| - | |Image:7.8.08gel.jpg | + | |[[Image:7.8.08gel.jpg|thumb|left|600px]] |
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Revision as of 21:51, 8 July 2008
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| 1. Simulations: Finish typing into designgui the simulations/rxn network. NOT USING SYNBIOSS. |
| 2. Restart/Redo PCR: Problem with original PCR was an error in the calculation of DNA concentration of product. 07-07-2008 we added only 1.0uL of PCR to digest reactions meaning only 3ng of DNA was in the solution. That was not a substantial amount of DNA to be seen in a electrophoretic gel. FIX - recalculated DNA concentration and is 3ng/1uL instead of 300ng/uL. Redid PCR2 with 100.0ng DNA instead of 1ng of DNA by adding 10.0uL of DNA. Will run gel with new PCR undigested (original PCR was digested). |
| 3. PCR: New PCR completed successfully. The RBS has been incorporated into the lambda cI gene, and the resulting DNA has a concentration of 23 ng/ul. Analysis by agarose gel electrophoresis confirmed that the DNA fragment is of the appropriate size (~780 bp). Top lane is 1kb ladder, third lane is PCR product. |
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