qPCR
@Zerina Rahic
General advantages of qPCR
Q‐PCR approaches combine the detection of target template with quantification by recording the amplification of a PCR product via a corresponding increase in the fluorescent signal associated with product formation during each cycle in the PCR.
- Provides fast and high-throughput detection and quantification of target DNA sequences in different matrices.
The lower time of amplification is facilitated by the simultaneous amplification and visualization of newly formed DNA amplicons.
- qPCR is safer in terms of avoiding cross contaminations because no further manipulation with samples is required after the amplification.
- qPCR include a wide dynamic range for quantification (7–8 Log10) and the multiplexing of amplification of several targets into a single reaction (Klein, 2002).
The multiplexing option is essential for detection and quantification in diagnostic qPCR assays that rely on the inclusion of internal amplification controls
- q‐PCR that uses fluorescence‐based detection offers greater sensitivity and enables discrimination of gene numbers across a wider dynamic range than is found with end‐point PCR
- PCR has inherent limitations, particularly those that result in biases in the template to product ratios of target sequences amplified during PCR from environmental DNA, with such amplification biases found to increase with increasing numbers of PCR cycles.
qPCR in eDNA
References