Label-free microarray imaging for direct detection of DNA hybridization and single-nucleotide mismatches.
Label-free microarray imaging for direct detection of DNA hybridization and single-nucleotide mismatches.
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DOI：
10.1016/j.bios.2009.12.032
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发表时间：
2010-03-15
影响因子：
12.6
通讯作者：
Unlü MS
中科院分区：
文献类型：
--
作者：
Ozkumur E;Ahn S;Yalçin A;Lopez CA;Cevik E;Irani RJ;DeLisi C;Chiari M;Unlü MS
提出了一种直接检测微阵列DNA杂交的新方法。光学干涉测量法用于无标记传感生物分子在玻璃表面的堆积，从而能够动态检测相互作用。通过对寡核苷酸固相杂交的高通量检测，证明了该方法的有效性。通过比较杂交前后获得的无标记微阵列图像，检测表面固定化探针与20个碱基对长的目标寡核苷酸的杂交。通过用低离子浓度缓冲液洗涤样品变性过程中的动态数据采集，将单核苷酸错配的双链熔融与高可信区间(>；97%)的完全匹配的双链区分开来。该技术简单、可靠、准确，不需要使用标记或二次试剂来监测寡核苷酸杂交。
A novel method is proposed for direct detection of DNA hybridization on microarrays. Optical interferometry is used for label-free sensing of biomolecular accumulation on glass surfaces, enabling dynamic detection of interactions. Capabilities of the presented method are demonstrated by high-throughput sensing of solid-phase hybridization of oligonucleotides. Hybridization of surface immobilized probes with 20 base pair-long target oligonucleotides was detected by comparing the label-free microarray images taken before and after hybridization. Through dynamic data acquisition during denaturation by washing the sample with low ionic concentration buffer, melting of duplexes with a single nucleotide mismatch was distinguished from perfectly matching duplexes with high confidence interval (>97%). The presented technique is simple, robust, and accurate, and eliminates the need of using labels or secondary reagents to monitor the oligonucleotide hybridization.
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影响因子：
14.9
作者：
Peterson, AW;Heaton, RJ;Georgiadis, RM
通讯作者：
Georgiadis, RM
影响因子：
8
作者：
Fishback, Price V.;Haines, Michael R.;Kantor, Shawn
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Kantor, Shawn
影响因子：
56.9
作者：
Golub, TR;Slonim, DK;Lander, ES
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Lander, ES
影响因子：
9.3
作者：
Koehne, JE;Chen, H;Li, J
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Li, J
影响因子：
5.4
作者：
Ramachandran, N;Larson, DN;LaBaer, J
通讯作者：
LaBaer, J