ABSTRACT
The targeting of specific DNA segments by molecular hybridization or DNA amplification by the polymerase chain reaction can be used to study variant or polymorphic sites in the genome. Preliminary studies showed that fluorescently labeled groups affixed to the very short oligonucleotide primers still allowed efficient amplification, but simplified the fingerprints. While it is obvious that many parameters of the amplification reaction can condition amplicon production as much as primer sequence, adequate resolution of amplification fragments will not only reveal the true number of amplified sites but ensure that DNA patterns obtained are reproducible. Although DNA amplification is most likely a reaction process departing from equilibrium, it may be argued that there are two reaction components: a standard Michaelis-Menten kinetic process discriminating target cognate DNA from noncognate competitor sites, followed by a virtually irreversible process that is thermodynamically driven by the cleavage of diphosphate from deoxynucleoside triphosphates.
