840:153g:Projects/project19/2011/10/13

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Today we tested the results of our PCR amplification by running the contents of each PCR tube as well as a DNA ladder (100bp - 3kbp) on a 1.0% agarose gel electrophoresis apparatus at 150V for 45 minutes. UV imagery of the results were only partially helpful as it appeared that additional time in the electric field would benefit interpretation of the size of our amplified product. We expect our gene fragment to be about 1650 bp. However, becuase we relied on cDNA compiled from the mRNA sequence, we cannot be sure that the gene is free from non-coding squences known as introns. If the amplified fragment is longer, a sequencing step will be required to find and then later excise the intron. In any case, we have learned that annealing temperatures of 42-47ÂșC produced the strongest results. Bands were visible, and our group was satiated for the week...

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