Abstract
The advent of recombinant DNA methodology, and most recently the introduction of the Polymerase Chain Reaction (PCR), have together made possible the characterization of a wide variety of different gene lesions underlying human genetic disease [9,18]. However, the major constraint on our ability to detect mutations in those cases where the gene responsible is already known, has often been the great size and complexity of the genes under study. One good example is provided by the gene encoding factor VIII (FVIII), the clotting factor deficient in haemophilia A; the 9kb FVIII mRNA is encoded by a gene comprising 26 exons spanning 186kb of the human X-chromosome [12]. We describe here two different PCR-based approaches designed to facilitate mutation detection and analysis; (i) a “directed search” strategy for screening potentially hypermutable sites and (ii) the use of ectopically-expressed mRNA transcripts as analytical material.
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Berg, LP. et al. (1991). Application of PCR to the Detection and Analysis of Point Mutations in the Human Factor VIII Gene. In: Rolfs, A., Schumacher, H.C., Marx, P. (eds) PCR Topics. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-75924-6_4
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DOI: https://doi.org/10.1007/978-3-642-75924-6_4
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