Locus-specific PCR primer sets previously characterised by Noller et al., (2003) [ 12] were used to amplify each of seven TR regions using single primer-paired reactions for all fifty-four isolanalysedlysed.
Long-range PCR was used to amplify ∼9.5 kb region of the mitochondrial genome across the major arc using a single primer set corresponding to nucleotides 6378 15896 (GenBank Accession number: NC_012920.1).
The objective of our study was thus to develop an innovative and cost-effective RT-qPCR assay, to simultaneously quantify the expression of five major cytokines involved in Th1/Th2 responses, IL-4, IL-10, IL-12B, IFN-γ and TNF-α, using a single primer set and hybridization temperature for three species of ruminants, cattle, sheep and goats, in a single assay.
To amplify the entire exon 2 of the DQB in the Japanese black bear, we initially used a single primer set URDQBex2-F1 and URDQBex2-R1 [ 39].
Because these analyses use the data obtained with single primer sets, their results are comparable with those from the majority of published reports.
Secondly, the amplification of a multi-gene family is necessarily problematic since not all loci and not all alleles at a locus will be detected using a single primers set.
The use of a single primer set can minimize the amplification bias that occurs by differences in Tm of PCR primers when using exon-specific multiplex primers.
Our multiplex amplification regime increased the detection as compared to any single primer set indicating the usefulness of using multiple primer sets in initial amplification of target genes.
Due to the presence of 11 different oligonucleotide sets for each specific gene being used on the microarray versus only a single primer set for a gene in a quantative PCR, the microarray is more effective at eliminating false positive or negative data and provides a more robust quantitation of changes in gene expression.
Due to the presence of 11 different oligonucleotide sets for each specific gene being used on the microarray versus only a single primer set for a gene in a quantitative PCR, the microarray is more effective at eliminating false positive or negative data and provides a more robust quantification of changes in gene expression.
