You’ll need to keep in mind that the length and composition of primers directly affects the PCR annealing temperature (Ta). A melting temperature (Tm) of 52°C to 58°C is a good starting range when designing primers. (Longer strands have higher melting temperatures, as do sequences ...
RT-PCR was used for the detection of RVA RNA by using primers NSP3F and NSP3R and a QuantiTect SYBR green RT-PCR kit. The following conditions were applied: incubation for 20 min at 50°C for the reverse transcription reaction and a preheating step at 95°C for 15 min for initial ...
Full-length and mRNA sequences for these reference genes were determined using the Paper Mulberry genome annotation file, and Primer Premier 5.0 was used for primer design with the following constraints: primer length, 18–24 bp; G.C. content, 45–55%; melting temperature, ~60 °C; and ...
The PCR program comprised initial denaturation at 94 °C for 4 min, 40 cycles of denaturation at 95 °C for 15 s and annealing at 56 °C for 10 s followed by melting curve analysis. PCR reactions with the universal bacterial primer pair HDA1/HDA2 [21], aimed at revealing the presence...
Agricultural, Aquatic and Forest Ecosystems: Destabilizing AgentsAssessing Insect Pollinator Populations Using Molecular Methods—Approaches and Metrics on Species Diversity, Abundance and Genetic DiversityAugmentative Biological Control Using Parasitoids for Fruit Fly ManagementBark and Wood-Boring Insects: Past ...
The PCR program comprised initial denaturation at 94 ◦C for 4 min, 40 cycles of denaturation at 95 ◦C for 15 s and annealing at 56 ◦C for 10 s followed by melting curve analysis. PCR reactions with the universal bacterial primer pair HDA1/HDA2 [21], aimed at revealing the ...
RTqPCR was performed for gene expression analyses using the QuantiNova™ SYBR Green PCR Kit (Qiagen, Hilden, Germany) following the manufacturer’s instructions. Briefly, master mix prepared on ice contained forward and reversed primers (test concentration each 0.5 µM) and cDNA or Yellow ...