HIGH-SPEED POLYMERASE CHAIN-REACTION IN CONSTANT FLOW
HIGH-SPEED POLYMERASE CHAIN-REACTION IN CONSTANT FLOW
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DOI：
10.1271/bbb.58.349
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发表时间：
1994-02-01
影响因子：
1.6
通讯作者：
YAMANE, T
中科院分区：
文献类型：
--
作者：
NAKANO, H;MATSUDA, K;YAMANE, T
介绍了一种新型的管式聚合酶链式反应器。使用薄的Teflon毛细管作为管式反应器，其中PCR的反应混合物由泵以恒定流速驱动。用三个连续的热阶段处理样品，用于DNA和引物的变性、退火和延伸，作为管中位置的函数。扩增产率约为商业热循环仪的一半。此外，总反应时间为12至18分钟，这是使用金属块的常规热循环仪通常所需时间的十分之一，确保了DNA片段的大量扩增。此外，该反应器还可用于快速循环序列。这种新的设备将很容易被纳入自动化和快速DNA分析系统的DNA测序。
A new simple reactor of the tubing type was developed for polymerase chain reaction (PCR). A thin Teflon capillary tube was used as a tubing reactor in which the reaction mixture of PCR was driven by a pump at a constant flow rate. The sample was treated with three successive thermal stages for denaturation, annealing, and elongation of DNA and primers as a function of the position in the tube. The amplification yield was about a half of that obtained by a commercial thermocycler. Moreover, the total reaction time from 12 to 18 min, which was one-tenth of the time generally required by conventional thermocyclers using metal blocks, assured substantial amplification of a DNA fragment. In addition, this reactor could be also used far rapid cycle-sequences. This new device will be easily incorporated into automated and rapid DNA analysis systems for DNA sequencing.