Evaluating digital PCR for the quantification of human nuclear DNA: determining target strandedness.

ABSTRACT: Digital polymerase chain reaction (dPCR) methodology has been asserted to be a "potentially primary" analytical approach for assigning DNA concentration. The essence of dPCR measurements is the independent dispersal of fragments into multiple reaction partitions, amplifying fragments containing a target nucleotide sequence until the signal from all partitions containing at least one such fragment rises above threshold, and then determining the proportion of partitions with an above-threshold signal. Should originally double-stranded DNA (dsDNA) fragments be converted into two single strands (ssDNA) prior to dispersal, the dPCR measurements could be biased high by as much as a factor of two. Realizing dPCR's metrological potential therefore requires analytical methods for determining the pr

SUBMITTER: Kline MC 

PROVIDER: S-EPMC7931668 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature