I already tested three different sequences and had problems with dimers and melt curve (low specificity). Any reliable primers? I'm studying rat microbiota with qPCR.
Marilia Cavalcanti Some short 16S-homologs sequences are present in bacterial genomes, and you can get non-target amplification for microbiota samples. Try to use several group-specific primers and analyse group-specific dynamics
i was thiniking that you use 16S rRNA gene primers that consists of both conserved and variable regions , try to use primers just for the V3 conserved region in 16S rRNA gene.