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采用变性方法通过免聚合酶链式反应(PCR)的压电传感检测基因组DNA片段化

Detection of fragmented genomic DNA by PCR-free piezoelectric sensing using a denaturation approach.

作者信息

Minunni Maria, Tombelli Sara, Fonti Jessica, Spiriti Maria M, Mascini Marco, Bogani Patrizia, Buiatti Marcello

机构信息

Dipartimento di Chimica, Università degli Studi di Firenze, 50019 Sesto Fiorentino, Italy.

出版信息

J Am Chem Soc. 2005 Jun 8;127(22):7966-7. doi: 10.1021/ja051345q.

Abstract

Label-free and real-time DNA sequence detection in PCR-amplified DNA samples can now be achieved by different approaches. On the contrary, only few works have been reported dealing with direct sequence detection in nonamplified genomic DNA. Here, a piezoelectric biosensor for direct detection of sequences in nonamplified genomic DNA is described. The system relies on real-time and label-free detection of the hybridization reaction between an immobilized probe and the complementary sequence in solution. The DNA probe is immobilized on the sensing surface (10 MHz quartz crystals), while the complementary sequence is present in the genomic DNA, previously fragmented with restriction enzymes.

摘要

现在,可以通过不同方法在PCR扩增的DNA样本中实现无标记实时DNA序列检测。相反,关于非扩增基因组DNA直接序列检测的报道较少。在此,描述了一种用于直接检测非扩增基因组DNA中序列的压电生物传感器。该系统依赖于固定化探针与溶液中互补序列之间杂交反应的实时无标记检测。DNA探针固定在传感表面(10 MHz石英晶体)上,而互补序列存在于先前用限制性酶切割的基因组DNA中。

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