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New PCR Primers for Human Diseases

Figure 1

Figure 2

Each primer set was used in PCR amplifications of DNA templates from infected serum samples, with genomic DNA from infected cells or with cDNA made from RNA isolated from infected cells. The control reactions were performed with DNA from both uninfected sera and human cells. The specificity of each primer set is shown in figure 1. Each amplification yielded a single product of the appropriate size in reactions with DNA from infected serum samples (figure 1 and figure 2), whereas amplification products were not detected in any of the negative controls, uninfected serum samples (figure 1).

Assay for the Factor V Mutation

The factor V (Leiden) mutation is associated with thrombophilia. In order to test for a point mutation of G1691A, the gene coding for human coagulation factor V, the factor V (Leiden) mutation assay is used. This assay requires an initial PCR amplification with subsequent digestion with the Mnl I restriction enzyme that yields a restriction fragment length polymorphism for the wild-type, heterozygous and homozygous genotypes (figure 2). Stratagenes primer set for performing the factor V mutation assay includes the Mnl I enzyme and the corresponding
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