用于材料科学和分析科学的多毫克量纯单链DNA样品的酶法制备。
Enzymatic preparation of multimilligram amounts of pure single-stranded DNA samples for material and analytical sciences.
作者信息
Nelissen Frank H T, Goossens Elles P M, Tessari Marco, Heus Hans A
机构信息
Department of Biophysical Chemistry, Institute for Molecules and Materials, Radboud University Nijmegen, 6525 AJ Nijmegen, The Netherlands.
Department of Biophysical Chemistry, Institute for Molecules and Materials, Radboud University Nijmegen, 6525 AJ Nijmegen, The Netherlands.
出版信息
Anal Biochem. 2015 Apr 15;475:68-73. doi: 10.1016/j.ab.2015.01.014. Epub 2015 Jan 29.
We present a method for high-yield production of multimilligram amounts of pure single-stranded DNA employing rolling circle amplification (RCA) and processing by restriction enzymes. Pure and homogeneous samples are produced with minimal handling time, reagents, and waste products. The RCA method is more than twice as efficient in dNTP incorporation than conventional polymerase chain reaction in producing end product. The validity and utility of the method are demonstrated in the production of a uniformly (13)C/(15)N-labeled 38-nt cocaine aptamer DNA used in nanosensing devices.
我们提出了一种通过滚环扩增（RCA）和限制性酶处理来高产多毫克量纯单链DNA的方法。该方法能够以最少的操作时间、试剂和废物产生纯净且均一的样品。在产生终产物方面，RCA方法在dNTP掺入上的效率是传统聚合酶链反应的两倍多。该方法的有效性和实用性在用于纳米传感装置的均匀（13）C/（15）N标记的38个核苷酸的可卡因适体DNA的生产中得到了证明。
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