At this time DNA has been extracted from 60+ blood samples and tested using one designed set of primers. The gels are displayed (40-50K) with short descriptions below. For an explanation of the setup and running of these gels see here (gel running).
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This first group of samples is shown with a control lane (on the far right side). This "master mix" contains all of the components necessary for the PCR reaction except the template (iguana) DNA. (The components include: filter purified distilled water, a packaged prepared PCR buffer, MgCl2, dNTPs [the building blocks of the DNA - A, T, G, C], the two primers, and Taq polymerase.) The control was run to be sure that no artifact DNA bands were being produced.
The bands of interest are located near the middle of the gel.
Notice that several of the "lanes" do not contain any bands. The
most obvious correlation between whether a lane has a band (or bands) of
DNA with these primers is the quality of the stock DNA. Samples with
low concentrations of do not produce bands. Many of the collected
blood samples have not provided high quality DNA (see also DNA
extraction methods tested).
From this point on, all of the samples have been tested at a 64o C annealing temperature. (I figured it was better to get some bands, even if they are not as specific as they could be, than to not know whether the primers will work at all with the extracted DNA for any one individual.) That is why these gels have more "stutter" bands (lighter bands above or below the primary bands).
For several of the individuals (male 35, male 121, & female 6) there
appear to be 2 distinct bands of equal intensity. These are potentially
individuals with 2 separate alleles (1 on each of 2 homologous chromosomes).
Using the primers designed for this microsatellite analysis the maximum
number of alleles present in each individual is 2 (since each of the 2
homologous chromosomes [1 from each parent] may contain a separate allele).
Each of the primers are designed to match only 1 location within the entire
genome, so only a single small section of the genome is copied/amplified
during the PCR process.
More . . .
More . . .
More . . .
And still more . . .
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