Real-time SNP analysis in secondary-structure-folded nucleic acids.
Real-time SNP analysis in secondary-structure-folded nucleic acids.
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DOI：
10.1002/anie.201004475
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发表时间：
2010-11-15
影响因子：
16.6
通讯作者：
Kolpashchikov, Dmitry M.
中科院分区：
文献类型：
--
作者：
Grimes, Jeffrey;Gerasimova, Yulia V.;Kolpashchikov, Dmitry M.
两条互补核酸链的杂交广泛用于实时PCR、DNA微阵列和活细胞中RNA监测技术中的特异性DNA/RNA序列分析。杂交探针的设计是基于AT和GC互补性，并且可能看起来很简单。然而，单链DNA和RNA分析物通常在测定条件下形成稳定的二级结构。这种折叠分析物的分析通常是复杂的，因为感兴趣的区域可能参与分子内杂交，并且变得无法与探针杂交。这种复杂性严重限制了灵敏度，并为检测两种分析物之间的单核苷酸差异造成了不可逾越的障碍。[1]这项研究展示了一种方法，允许在室温下真实的时间内分析折叠分析物中的单核苷酸多态性（SNP）。SNP代表了人类最丰富的遗传变异类别，占两个个体基因组之间差异的80-90%。[2]有超过300万个经过验证的SNP，尽管实际数量被认为在1000万的范围内。SNP分析在基于群体的遗传风险评估、分子诊断、药物开发、连锁分析和法医应用中的身份测试中是重要的。[3]第一章
Hybridization of two complementary nucleic acid strands is extensively used in the analysis of specific DNA/RNA sequences in real-time PCR, DNA microarrays and the techniques for RNA monitoring in living cells. The design of the hybridization probes is based on AT and GC complementarity and may seem straightforward. However, single-stranded DNA and RNA analytes often form stable secondary structures under assay conditions. The analysis of such folded analytes is often complicated since a region of interest may be involved in intramolecular hybridization and become inaccessible for hybridization with a probe. This complication severely limits sensitivity and creates and insurmountable obstacle for the detection of single nucleotide differences between two analytes.[1] This study demonstrates an approach that allows analysis of single nucleotide polymorphisms (SNPs) in folded analytes in real time at room temperature.SNPs represent the most abundant class of genetic variations in humans, accounting for 80–90% of the difference between the genomes of two individuals.[2] There are over 3 million validated SNPs, although the actual number is believed to be in the range of 10 million. SNP analysis is important in population-based genetic risk assessment, molecular diagnostics, pharmaceutical drug development, linkage analysis, and identity testing in forensic applications.[3]
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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：
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：
Kushon, SA;Jordan, JP;Armitage, BA
通讯作者：
Armitage, BA
影响因子：
56.9
作者：
Gabriel, SB;Schaffner, SF;Altshuler, D
通讯作者：
Altshuler, D