Standardized determination of real-time PCR efficiency from a single reaction set-up

Standardized determination of real-time PCR efficiency from a single reaction set-up.

We propose a computing method for the estimation of real-time PCR amplification efficiency. It is based on a statistic delimitation of the beginning of exponentially behaving observations in real-time PCR kinetics. PCR ground fluorescence phase, non-exponential and plateau phase were excluded from the calculation process by separate mathematical algorithms. We validated the method on experiment...

Quantitative Real-time Polymerase Chain Reaction (qRT-PCR). Fatemeh Mehrpooyan*

Evaluation of a new set of Real-Time PCR for Brucella detection within human and animal samples

A quantitative TaqMn Real-Time PCR assay was developed and its diagnostic value on human serum and livestock samples were evaluated. Brucella species could be distributed through communities as a biological agent. Rapid detection of biological threat agents is critical for timely therapeutic administration. Quantitative real-time PCR provides a rapid, sensitive and specific tool for molecular i...

Standard Addition Quantitative Real-Time PCR (SAQPCR): A Novel Approach for Determination of Transgene Copy Number Avoiding PCR Efficiency Estimation

Quantitative real-time polymerase chain reaction (qPCR) has been previously applied to estimate transgene copy number in transgenic plants. However, the results can be erroneous owing to inaccurate estimation of PCR efficiency. Here, a novel qPCR approach, named standard addition qPCR (SAQPCR), was devised to accurately determine transgene copy number without the necessity of obtaining PCR effi...

Species Specific DNA Profiling Mycobacterial Genomes Using Polymerase Chain Reaction with Single Universal Primer (UP-PCR)

Three tuberculous, twenty-one non-tuberculous mycobacterial (NTM) reference strains and seventy two isolates classified by biochemical tests were shown to produce specific sets of DNA fragments in a polymerase chain reaction with single universal primer (UP-PCR). A rather wide limit of tolerance for variations in procedure of PCR mixture preparation and thermocycling parameters was found. There...