Minimizing the time required for DNA amplification by efficient heat transfer to small samples.
Hot-air temperature cycling of 1- to 10-microliters samples in glass capillary tubes can amplify DNA by the polymerase chain reaction in 15 min or less. A rapid temperature cycler of low thermal mass was constructed to change sample temperatures among denaturation, annealing, and elongation segments in a few seconds. After 30 cycles of 30 s each, a 536-bp beta-globin fragment of human genomic DNA was easily visualized with ethidium bromide on agarose gels. With rapid cycling, amplification yield depended on polymerase concentration. The time required for DNA amplification can be markedly reduced from prevailing protocols if appropriate equipment and sample containers are used for rapid heat transfer to the sample.
Wittwer
C T
CT
Department of Pathology, University of Utah Medical School, Salt Lake City 84132.
Fillmore
G C
GC
Garling
D J
DJ
D004247:DNA / Q000378:metabolism; D005784:Gene Amplification; D006358:Hot Temperature; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D013997:Time Factors
D004247:DNA