Physical Characteristics of Six New Thermocyclers

[摘要] Since the publication of the first article describing PCR, thermocyclers have become a staple in academic and industrial laboratories (1). The thermocycler is a programmable cycling incubator that performs repeated PCR steps of DNA denaturation, primer annealing, and primer elongation at defined intervals. Rapid heat transfer from the heating block to the in-tube sample liquid ensures a high efficiency of amplicon multiplication; therefore, a thermal processor should guarantee temperature uniformity for all samples within an individual run as well as run-to-run repeatability.PCR-based protocols can give unsatisfying results (2)(3). Several collaborative studies have shown weak reproducibility with random amplified polymorphic DNA (RAPD) protocols (4)(5). One reason might be the influence of the thermocycler on amplification efficiency.Despite its striking importance for PCR, the literature on thermocyclers is scarce. Some studies were published on the first generation of cyclers (6)(7)(8). Others determined the amplification efficiency but did not evaluate the physical characteristics (9)(10). The impact on PCR of the variation within thermocyclers, with regard to their thermocycling settings, has not been fully determined. The goal of this study was to define the physical characteristics of performance of the latest generation of thermocyclers and to discuss the influence of the physical properties on amplification efficiency.Six new thermocyclers were selected for this performance study: ( A ) Gene Amp 9700 (Applied Biosystems), ( B ) Multicycler PTC 200 (MJ Research, Inc.), …

[学科分类] 过敏症与临床免疫学