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.
In recent years there has been great interest in quantitative polymerase chain reaction. Consequently, a large number of assays have been developed, of which the one using non-homologous competitors is arguably the most precise. Despite widespread applications, currently there is no simple method to synthesize such competitors. Here a facile and cost-effective, single-step method for synthesis of recombinant, non-homologous, competitor complementary DNA is described. The method can be adapted to generate competitors of any size and sequence. The entire procedure is quick, straightforward and does not require any specialized equipment except a standard thermocycler.
Phulwinder
K Grover
KG
Department of Surgery, Flinders University School of Medicine, Flinders Medical Centre, South Australia.
D000818:Animals; D017931:DNA Primers / Q000138:chemical synthesis* / Q000737:chemistry; D018076:DNA, Complementary / Q000138:chemical synthesis* / Q000737:chemistry; D008297:Male; D009692:Nucleic Acid Heteroduplexes / Q000138:chemical synthesis / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D011516:Prothrombin / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar
D017931:DNA Primers; D018076:DNA, Complementary; D009692:Nucleic Acid Heteroduplexes; D011516:Prothrombin