Loss of Heterozygosity Studies Revisited

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Presentation transcript:

Loss of Heterozygosity Studies Revisited Kathryn Farrand, Lydija Jovanovic, Brett Delahunt, Bryan McIver, Ian D. Hay, Norman L. Eberhardt, Stefan K.G. Grebe The Journal of Molecular Diagnostics Volume 4, Issue 3, Pages 150-158 (August 2002) DOI: 10.1016/S1525-1578(10)60696-4 Copyright © 2002 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 GeneScan electropherograms of three duplicate microsatellite PCR (D17S695) of the same, 100% amplifiable reference DNA sample at template inputs of 600 pg (top), 240 pg (middle) and 42 pg (bottom). Alleles are indicated by shading, other peaks represent stutter and a-tails. The abscissas of each panel show the allele sizes in bp, the ordinates show the allele peak heights in arbitrary fluorescence units. Allele peak heights fall slightly with decreasing template input, but remain satisfactory in all three experiments. However, only the 600 pg template input samples display no differences in allele height between the two duplicate samples. At 240 pg DNA input, significant allelic imbalance is evident with the peak height of the shorter allele in duplicate 2 being decreased. At 42 pg template input, a complete allelic drop-out of the longer allele of duplicate 1 is observed. The Journal of Molecular Diagnostics 2002 4, 150-158DOI: (10.1016/S1525-1578(10)60696-4) Copyright © 2002 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Figure 2 shows the data analysis screen of a typical LightCycler experiment to determine the amplifiable DNA content of thyroid tumor samples. The uppermost portion shows the analysis settings. The table on the left, below the settings, contains the input DNA standards (10 ng, 4 ng, 400 pg, and 40 pg of 100% amplifiable reference DNA), the calculated amplifiable DNA content of the standards and of 10 unknown samples (based on the generated standard curve) and the crossing points (PCR cycle number when the exponential phase was reached) for all standards and samples. The last row contains a negative control sample. The actual PCR fluorescence profiles for the standards and samples are depicted in the panel to the right of the table. Beneath the fluorescence profiles, the standard curve and its associated parameters are shown. The Journal of Molecular Diagnostics 2002 4, 150-158DOI: (10.1016/S1525-1578(10)60696-4) Copyright © 2002 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions