Abstract
Chimerism analysis is a valuable tool in medical diagnostics, population genetics, and food control. The selective quantification of one allele in samples containing higher amounts of very similar sequences can be achieved by several different techniques. Immunoglobulin and T-cell receptor genes, Y-chromosomal sequences [1], restriction fragment length polymorphisms (RFLP), short tandem repeats (STR), and variable number of tandem repeats (VNTR) [2] are chosen as typical targets that have been selectively quantified in PCR-based chimerism analysis. Genetic length variations and RFLPs [3] can be identified by fragment length, but quantification must be performed after electrophoretic separation of the PCR products, e.g., by capillary electrophoresis [4]. Unfortunately, the dynamic range of this time consuming method is approximately 1:20, i.e., not less than one copy of one allele among 20 copies of another. The highly underpopulated allele is not amplified to detectable amounts before the plateau phase of the PCR is reached.
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Reuter, H., Tews, B., Wilhelm, J., Hahn, M. (2004). Quantitative Chimerism Analysis by Allele-Specific Real-Time PCR of a 10 bp Insertion/Deletion Polymorphism within the Promotor Region of Factor Vllc. In: Wittwer, C., Hahn, M., Kaul, K. (eds) Rapid Cycle Real-Time PCR — Methods and Applications. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-18840-4_19
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DOI: https://doi.org/10.1007/978-3-642-18840-4_19
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