A comparison of hybridization efficiency between flat glass and channel glass solid supports

被引:4
|
作者
Betanzos-Cabrera, Gabriel [1 ]
Harker, Brent W. [2 ]
Doktycz, Mitchel J. [3 ]
Weber, James L. [4 ]
Beattie, Kenneth L. [3 ]
机构
[1] Univ Autonoma Estado Hidalgo, Inst Ciencias Salud, Area Acad Nutr, Hidalgo 42000, Mexico
[2] Univ Notre Dame, Ctr Global Hlth & Infect Dis, Dept Biol Sci, Notre Dame, IN 46556 USA
[3] Oak Ridge Natl Lab, Biosci Div, Oak Ridge, TN 37831 USA
[4] Marshfield Med Res, Ctr Genet Med, Marsfield, WI USA
关键词
channel glass; flat glass; oligonucleotide arrays; capture probes; hybridization; sensitivity;
D O I
10.1007/s12033-007-9001-z
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Two different solid supports, channel glass and flat glass, were compared for their affect on the sensitivity and efficiency of DNA hybridization reactions. Both solid supports were tested using a set of arrayed, synthetic oligonucleotides that are designed to detect short insertion/deletion polymorphisms (SIDPs). A total of 13 different human SIDPs were chosen for analysis. Capture probes, designed for this test set, were covalently immobilized on substrates. Hybridization efficiency was assessed using fluorescently labeled stacking probes which were preannealed to the target and then hybridized to the support-bound oligonucleotide array; the hybridization pattern was detected by fluorescence imaging. It was found that structural features of nucleic acid capture probes tethered to a solid support and the molecular basis of their interaction with targets in solution have direct implications on the hybridization process. Our results demonstrate that channel glass has a number of practical advantages over flat glass including higher sensitivity and a faster hybridization rate.
引用
收藏
页码:71 / 80
页数:10
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