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The Polymerase Chain Reaction: - Essay Example

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If your DNA has migrated to the band designated at 100 bp position, then your genotype is homozygous for the absence of TPA-25 gene Alu insertion polymorphism (-/- or short/short). However if it migrates to the 400 bp band position you are considered homozygous for the presence…
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The Polymerase Chain Reaction:
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Download file to see previous pages At this temperature the hydrogen bonds holding the two strands of the double helix are broken and the DNA separates into single strands.
The 50C step lets the primers anneal to the complementary sequences specific to them in the template DNA. The temperature for this step is carefully chosen so as to allow the primers to bind to the correct complements. The exact annealing temperature depends on the length and sequence of the primers used for the procedure. A lower annealing temperature might result in amplification of non-specific sequences due to incorrect binding of primers.
72C is the temperature at which the Taq DNA polymerase is most active. At this temperature the polymerase binds and adds nucleotides to the 3′ ends of annealed primers resulting in the extension of complementary DNA.
Under ideal conditions, the amount of PCR products doubles at each cycle and the primer directed DNA extension amplify the region between the primers in a geometric manner. Hence after 30 cycles it shows a 2^30 amplification which theoretically is an amplification factor of a billion approximately.
5). Calculate the allelic frequencies for the long and short alleles. (This is simply means the proportion of the TPA-25 gene that is in the long form in the population studied but NB each homozygote has two copies of that particular allele. For example if the data gave 25 long/long; 32 long/short and 43 short/short, then the frequency of the long allele is (25 x 2) + 32 (Total number of long alleles) divided by the Total number of alleles (long and short) i.e. 200.
6). In this case the polymorphism is harmless but for other genes, which can be screened just as easily, this may not be the case. Certain polymorphisms are indicative of certain genetic diseases and PCR provides a very effective way of screening in many cases.
Suppose that in this case the insertion of the Alu element disrupts the function of the gene and prevents a ...Download file to see next pagesRead More
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