The specificity of the primer was assessed by submitting its sequence to the GenBank Basic Local Alignment Search Tool (BLAST) algorithm, and was also tested with DNA extracted from several ascomycetous, basidiomycetous, and zygomycetous taxa.
RT-qPCR analysis used relative quantification with the amplification efficiency of each primer pair assessed by serial dilutions of the cDNA pool.
These primer sets assessed 4 CpG dinucleotides spanning nucleotides +184 289 (as the first ATG +1, GenBank accession number AF029081) in the CpG island.
Primers were assessed in silico to determine specificity of the primer set for a single genomic region and for the presence of SNPs in the targeted genomic region which could decrease primer binding and amplification efficiency.
The specificity of the primers was assessed with a panel of bacteria representing mastitis-negative control species.
The accuracy and specificity of the primers was assessed using genomic DNA extracted from meat samples of known sex.
The sensitivity and specificity of these primers were assessed by testing eight raw human sewage samples and 265 feces from 12 different species in Saskatchewan.
The sensitivity of the PCR primers was assessed with conventional and real-time PCR and serially diluted genomic DNA.
First, the primers were assessed using the probematch service from the Ribosomal Database Project [15].
The efficiency of the primers was assessed using a 10-fold dilution series of the recombinant plasmid and standard male genomic DNA.
