超声波降解DNA。

H I Elsner, E B Lindblad
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引用次数: 102

摘要

超声波对水溶液中的DNA和生物组织中的DNA的作用结果不同。在与临床应用的超声强度相当的情况下,超声能够降解水溶液中的纯化DNA,使超声成为体外制备DNA片段的有用工具。溶液中DNA的超声波降解是通过破坏氢键和DNA螺旋的单链和双链断裂来发生的。两种机制主要负责:空化和热或机械效应。在低强度超声下可见稳定的空化。当超声强度增加到2w /cm2以上时,由于瞬态空化产生自由基,单链断裂增加。超声处理后,所得DNA片段的分布接近100-500 bp的较低尺寸极限。DNA螺旋的断裂主要发生在氧原子和碳原子之间,导致DNA片段的5'端被磷酸化,3'端有游离醇。在降解DNA螺旋方面相对缺乏特异性,使得超声波作为限制性内切酶获得的高度特异性碎片的补充替代。
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Ultrasonic degradation of DNA.

Different results are obtained when DNA in aqueous solution and DNA in biological tissue are exposed to ultrasound. At intensities of ultrasound comparable to those applied clinically, ultrasonication is able to degrade purified DNA in aqueous solution, making ultrasonication a useful tool for preparing DNA fragments in vitro. Ultrasonic degradation of DNA in solution occurs by breaking hydrogen bonds and by single-strand and double-strand ruptures of the DNA helix. Two mechanisms are mainly responsible: cavitation and a thermal or mechanical effect. Stable cavitation is seen at low intensities of ultrasound. Increasing the intensity of the ultrasound above 2 W/cm2 is followed by increases in single-strand ruptures due to the creation of free radicals by transient cavitation. Following sonication, the distribution of the resulting DNA fragments approaches a lower size limit of 100-500 bp. Breaks in the DNA helix occur mainly between oxygen and carbon atoms, resulting in DNA fragments with a phosphorylated 5' end and a free alcohol at the 3' end. The relative lack of specificity in degrading the DNA helix makes ultrasonication a complementary alternative to the highly specific fragmentation obtained by restriction endonucleases.

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