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In recent years there has been great interest inquantitative polymerase chain reaction. Consequently, a large number of assayshave been developed, of which the one using nonhomologous competitors isarguably the most precise. Despite widespread applications, currently there isno simple method to synthesize such competitors. Here a facile andcosteffective, singlestep method for synthesis of recombinant, nonhomologous,competitor complementary DNA is described. The method can be adapted togenerate competitors of any size and sequence. The entire procedure is quick,straightforward and does not require any specialized equipment except astandard thermocycler.

biological chemistry

A Simple Single-Step Method for the Synthesis of     Recombinant Non-Homologous Competitor cDNA and Its Implications for     Quantitative PCR in Biological Sciences and Molecular     Diagnostics