(E) Melting curve analysis of the PCR products generated in (D). (F) The appearance of a second peak (arrow) at a lower-than-expected melting temperature strongly suggests primer–dimer or the formation of another nonspecific product. Moreover, the divergence of amplification curves between ...
However, the minor drawback of a multiplex SYBR green system is unable to predict the presence or absence of any particular gene or primer-dimer by specific Ct-value. Melting curve-based detection, wherein the dissociation points of three genes are distinct, and any flat curve on that ...
conc. hot-start reaction mix designed for amplification and detection of a specific DNA sequence (if suitable PCR primers are provided) followed by high resolu- tion melting curve anal 13、ysis for detection of sequence variants among several samples. A separate 25 mM MgCl2 stock solution, ...
Figure 1.The melting curve of the triple Eva Green real-time PCR assay developed in this study. Table 2.Primers that were used to differentiate EHP, IHHNV, and DIV1 in this study. 3.2. Determination of the Optimal Reaction Conditions ...