Utilization of Digital PCR in Quantity Verification of Plasmid Standards Used in Quantitative PCR.

ABSTRACT: Quantitative PCR (qPCR) is a widely used method for nucleic acid quantification of various pathogenic microorganisms. For absolute quantification of microbial load by qPCR, it is essential to create a calibration curve from accurately quantified quantification standards, from which the number of pathogens in a sample is derived. Spectrophotometric measurement of absorbance is a routine method for estimating nucleic acid concentration, however, it may be affected by presence of other potentially contaminating nucleic acids or proteins and salts. Therefore, absorbance measurement is not reliable for estimating the concentration of stock solutions of quantification standards, based on which they are subsequently diluted. In this study, we utilized digital PCR (dPCR) for absolute quantificatio

SUBMITTER: Beinhauerova M 

PROVIDER: S-EPMC7403525 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature