Determination of DNA melting temperatures in diffusion-generated chemical gradients

被引：12

作者：

Liedl, Tim

Simmel, Friedrich C.

机构：

[1] LMU Munich, Dept Phys, D-80539 Munich, Germany

[2] LMU Munich, Ctr Nanosci, D-80539 Munich, Germany

来源：

ANALYTICAL CHEMISTRY | 2007年 / 79卷 / 14期

关键词：

D O I：

10.1021/ac070242i

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

For fast and reliable determination of DNA melting temperatures with single-nucleotide resolution in a microfluidic setup, stable gradients of the denaturing agent formamide were generated by means of diffusion. Formamide lowers the melting temperature of DNA, and a given formamide concentration can be mapped to a corresponding virtual temperature along the formamide gradient. We applied this concept to determine the melting temperatures of five sets of dye- and quencher-labeled oligonucleotides of different lengths. Differences in the length of complementary sequences of only one nucleotide as well as a single nucleotide mismatch can be detected with this method. Comparison with conventional melting temperature measurements based on temperature scans yields very good agreement.

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收藏

页码：5212 / 5216

页数：5

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