FRugally Optimized DNA Octamer (FRODO): DNA Vector and Uses Thereof For Detecting HIV and SIV

Quantitative polymerase chain reactions (qPCRs) are commonly employed to enumerate genes of interest among particular biological samples. Insertion of PCR amplicons into plasmid DNA is a mainstay for creation of known quantities of target sequences to standardize quantitative PCRs. Typically, one amplicon is inserted into one plasmid construct, the plasmid is then amplified, purified, serially diluted, and then quantified to be used to enumerate target sequences in unknown samples. As qPCR is often used to detect multiple amplicons simultaneously, individual qPCR standards are often desired to be normalized one to another. Unlike prior methods using separate plasmid constructs for each target sequence, FRODO incorporates eight amplicons into one plasmid construct ensuring equivalent template copy numbers for all amplicons. Amplifying, purifying, diluting and quantifying one plasmid construct rather than eight individual constructs streamlines standard curve qPCR analyses, reducing reagents and simplifying normalization between amplicons.