Amplification of nonspecific products in quantitative polymerase chain reactions (qPCR).

ABSTRACT: Quantitative PCR allows the precise measurement of DNA concentrations and is generally considered to be straightforward and trouble free. However, a survey with 93 validated assays for genes in the Wnt-pathway showed that the amplification of nonspecific products occurs frequently and is unrelated to C_q or PCR efficiency values. Titration experiments showed that the occurrence of low and high melting temperature artifacts was shown to be determined by annealing temperature, primer concentration and cDNA input. To explore the range of input variations that occur in the normal use of the Cre assay these conditions were mimicked in a complete two-way design of template -plasmid DNA- and non-template -mouse cDNA- concentrations. These experiments showed that the frequency of the amp

SUBMITTER: Ruiz-Villalba A 

PROVIDER: S-EPMC5727009 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature