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使用多重化 CMOS 微阵列优化并比较合成或固定在各个电极上的寡核苷酸与 DNA 探针的结合。

Use of a multiplexed CMOS microarray to optimize and compare oligonucleotide binding to DNA probes synthesized or immobilized on individual electrodes.

机构信息

CombiMatrix Corporation, 6500 Harbour Heights Parkway, Suite #202, Mukilteo, WA 98275, USA.

出版信息

Sensors (Basel). 2010;10(8):7371-85. doi: 10.3390/s100807371. Epub 2010 Aug 5.

DOI:10.3390/s100807371
PMID:22163607
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3231185/
Abstract

The CombiMatrix microarray with 12,544 electrodes supports in situ electrochemical synthesis of user-defined DNA probes. As an alternative, we immobilized commercially synthesized DNA probes on individual electrodes coated with electropolymerized polypyrrole (Ppy). Hybridization was measured using a biotinylated target oligonucleotide and either Cy5-streptavidin and fluorescence detection or horseradish peroxidase-streptavidin and enzyme-enhanced electrochemical detection. Detection efficiencies were optimized by varying the deposition of the Ppy, the terminal groups on the DNA probes, and other factors that impacted fluorescence quenching and electrical conductivity. Optimized results were compared against those obtained using a microarray with the same DNA sequences synthesized in situ. Immobilized probes produced higher fluorescence signals, possibly by providing a greater stand off between the Cy5 on the target oligonucleotide and the quenching effects of the Ppy and the platinum electrode.

摘要

CombiMatrix 微阵列有 12544 个电极，支持用户定义的 DNA 探针的原位电化学合成。作为替代方法，我们将商业合成的 DNA 探针固定在涂有电聚合聚吡咯 (Ppy) 的单个电极上。使用生物素化的靶寡核苷酸和 Cy5-链霉亲和素和荧光检测或辣根过氧化物酶-链霉亲和素和酶增强电化学检测来测量杂交。通过改变 Ppy 的沉积、DNA 探针的末端基团和其他影响荧光猝灭和电导率的因素来优化检测效率。优化结果与使用原位合成相同 DNA 序列的微阵列获得的结果进行了比较。固定探针产生了更高的荧光信号，这可能是因为在靶寡核苷酸上的 Cy5 与 Ppy 和铂电极的猝灭效应之间提供了更大的间隔。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/3efa4c593576/sensors-10-07371f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/ab2808ca14a1/sensors-10-07371f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/13d97e6b6fec/sensors-10-07371f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/2a2a0d07be1a/sensors-10-07371f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/0d548a290065/sensors-10-07371f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/10823de41da8/sensors-10-07371f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/4c83b49e57f2/sensors-10-07371f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/6a7100aa5808/sensors-10-07371f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/3efa4c593576/sensors-10-07371f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/ab2808ca14a1/sensors-10-07371f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/13d97e6b6fec/sensors-10-07371f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/2a2a0d07be1a/sensors-10-07371f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/0d548a290065/sensors-10-07371f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/10823de41da8/sensors-10-07371f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/4c83b49e57f2/sensors-10-07371f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/6a7100aa5808/sensors-10-07371f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff00/3231185/3efa4c593576/sensors-10-07371f8.jpg

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