Classification of fowl adenoviruses by use of phylogenetic analysis and high-resolution melting-curve analysis
of the hexon L1 gene region.
was performed using 95[degrees]C for 60 s, 40[degrees]C for 20s, and heating to 80[degrees]C with a ramp rate of 0.
c] for fast COLD-PCR for a given amplicon is defined by first amplifying a wild-type sample via conventional PCR and then conducting a melting-curve analysis
(ramping at 0.
The presence of a KRAS mutation is detected either by an increase of fluorescence during PCR amplification or by post-PCR fluorescent melting-curve analysis
The amplification program was followed by a melting-curve analysis
consisting of 15-s holds at 95 [degrees]C, 45 [degrees]C, and 95 [degrees]C.
High-resolution melting-curve analysis
(HRM) is a simple and cost-effective post-PCR technique that can be used for methylation profiling (20).
During melting-curve analysis
, the progressive rise in temperature diminished the interaction between SGI and acceptor fluorophore as the duplex amplicons dissociated.
was performed by use of previously described methods (7) with LightScanner Software (version 1.
Rapid detection and differentiation of Newcastle disease virus by real-time PCR with melting-curve analysis
The accepted primer pairs showed no nonspecific or dimer amplification on an agarose gel and no nonspecific peaks in the temperature range of the specific cDNA and gDNA peaks in the melting-curve analysis
In contrast to traditional melting-curve analysis
, high-resolution melting with the HR-1 instrument reliably detects single-base differences in homozygous and heterozygous sequences (17).
In this assay, a 165-bp fragment of the prothrombin gene is amplified, and the different allelic variants are distinguished by melting-curve analysis
using the fluorescence resonance energy transfer principle.