Increased efficiency of genetic profiling through quantity and quality assessment of fluorescently labeled oligonucleotide primers.
Optimizing the amount of primer to use in PCR amplification is one of the most important steps when developing protocols for genetic profiling, where subtle changes in primer concentration result in major impacts on the amount of desired product that is amplified. However; there are frequently discrepancies between the reported and actual quantity of primers delivered by suppliers, resulting in a need for re-optimization of conditions between primer orders and limiting the ability to standardize conditions between laboratories. To increase the consistency of genetic profiling protocols, we have developed a simple method to assess the quantity and quality of fluorescently labeled primers and therefore standardize reaction conditions through time and across laboratories. The method is based on analysis by electrophoresis with an automated fluorescent DNA analyzer.
Frasier
Timothy R
TR
Trent University, Peterborough, ONT, Canada. timothyfrasier@trentu.ca
White
Bradley N
BN
D017931:DNA Primers / Q000378:metabolism* / Q000592:standards*; D005456:Fluorescent Dyes / Q000378:metabolism* / Q000592:standards*; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards*; D011786:Quality Control; D012015:Reference Standards
D017931:DNA Primers; D005456:Fluorescent Dyes