Fluorescence-based detection of short DNA sequences under non-denaturing conditions
Fluorescence-based detection of short DNA sequences under non-denaturing conditions
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DOI：
10.1016/j.bmc.2007.04.063
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发表时间：
2008-01-01
影响因子：
3.5
通讯作者：
Frank-Kamenetskii, Maxim D.
中科院分区：
文献类型：
--
作者：
Smolina, Irina V.;Kuhn, Heiko;Frank-Kamenetskii, Maxim D.
肽核酸（PNA）以高度序列特异性的方式打开双链DNA的能力使得在非变性条件下在基因组DNA的背景或内部检测短DNA序列成为可能。为此，双链DNA中选定的标记位点被一对PNA开启器局部打开，从而将目标区域（20- 30bp）内的一条链转化为单链形式。在这个可接近的DNA序列上组装了一个圆形寡核苷酸探针，它作为滚环扩增（RCA）的模板。均质和异质分析格式进行了研究，因为是基于荧光扩增子检测的不同格式。我们最近的数据与固定化分析表明，在质粒和细菌染色体DNA标记序列可以成功地检测。(C) 2007 Elsevier Ltd.版权所有。
The ability of peptide nucleic acid (PNA) to open up duplex DNA in a highly sequence-specific manner makes it possible to detect short DNA sequences on the background of or within genomic DNA under non-denaturing conditions. To do so, chosen marker sites in double-stranded DNA are locally opened by a pair of PNA openers, thus transforming one strand within the target region (20-30 bp) into the single-stranded form. Onto this accessible DNA sequence a circular oligonucleotide probe is assembled, which serves as a template for rolling circle amplification (RCA). Both homogeneous and heterogeneous assay formats are investigated, as are different formats for fluorescence-based amplicon detection. Our recent data with immobilized analytes suggest that marker sequences in plasmid and bacterial chromosomal DNA can be successfully detected. (C) 2007 Elsevier Ltd. All rights reserved.