A single molecular beacon probe is sufficient for the analysis of multiple nucleic acid sequences.
A single molecular beacon probe is sufficient for the analysis of multiple nucleic acid sequences.
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DOI：
10.1002/cbic.201000287
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发表时间：
2010-08-16
期刊：
影响因子：
3.2
通讯作者：
Kolpashchikov, Dmitry M.
中科院分区：
文献类型：
--
作者：
Gerasimova, Yulia V.;Hayson, Aaron;Ballantyne, Jack;Kolpashchikov, Dmitry M.
分子信标（MB）探针是双标记的发夹状寡脱氧核糖核苷酸，其广泛用于特异性RNA/DNA分析物的实时检测。在MB探针中，环片段与分析物互补：因此，分析每个新的分析物序列需要独特的探针。寡核苷酸与两种染料的缀合和随后的纯化程序增加了MB探针的成本，从而减少了它们在多重形式中的应用。在这里，我们演示了如何一个MB探针可以用于分析的任意核酸。该方法利用两个寡核苷酸衔接子链，每个衔接子链含有与分析物互补的片段和与MB探针互补的片段。分析物的存在导致MB探针与四分复合物中的两条DNA链缔合。MB探针荧光报告该复合物的形成。在该设计中，MB不直接结合分析物;因此，MB序列与分析物无关。在这项研究中，使用一种通用MB探针对三个人类多态性位点进行基因分型。这种方法有望降低多重实时检测的成本，提高单核苷酸多态性基因分型的准确性。
Molecular beacon (MB) probes are dual-labeled hairpin-shaped oligodeoxyribonucleotides that are extensively used for real-time detection of specific RNA/DNA analytes. In the MB probe, the loop fragment is complementary to the analyte: therefore, a unique probe is required for the analysis of each new analyte sequence. The conjugation of an oligonucleotide with two dyes and subsequent purification procedures add to the cost of MB probes, thus reducing their application in multiplex formats. Here we demonstrate how one MB probe can be used for the analysis of an arbitrary nucleic acid. The approach takes advantage of two oligonucleotide adaptor strands, each of which contains a fragment complementary to the analyte and a fragment complementary to an MB probe. The presence of the analyte leads to association of MB probe and the two DNA strands in quadripartite complex. The MB probe fluorescently reports the formation of this complex. In this design, the MB does not bind the analyte directly; therefore, the MB sequence is independent of the analyte. In this study one universal MB probe was used to genotype three human polymorphic sites. This approach promises to reduce the cost of multiplex real-time assays and improve the accuracy of single-nucleotide polymorphism genotyping.
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影响因子：
3.2
作者：
Gerasimova, Yulia V.;Cornett, Evan;Kolpashchikov, Dmitry M.
通讯作者：
Kolpashchikov, Dmitry M.
影响因子：
15
作者：
Du, H;Disney, MD;Krauss, TD
通讯作者：
Krauss, TD
影响因子：
15
作者：
Kolpashchikov, Dmitry M.
通讯作者：
Kolpashchikov, Dmitry M.
DOI：
10.1073/pnas.85.23.8790
发表时间：
1988-12-01
影响因子：
11.1
作者：
CARDULLO, RA;AGRAWAL, S;WOLF, DE
通讯作者：
WOLF, DE
影响因子：
15
作者：
Conlon, Patrick;Yang, Chaoyong James;Tan, Weihong
通讯作者：
Tan, Weihong