利用DNA序列方位性進行預雜合以增強微陣列探針-標的物之雜合效率

Enhancement of Target-DNA Hybridization Efficiency by Pre-hybridization on Sequence Orientated Microarrayed Probes

楊登凱(Deng-Kai Yang)

指導教授 ： 吳瑞璋

中原大學/工學院/化學工程學系/碩士(2008年)

https://doi.org/10.6840/cycu200800377

摘要

本研究於兩序列對調之微陣列探針預先雜交寡核苷酸片段（Oligo-nucleotide），探討其對於去氧核醣核苷酸（DNA）標的物（Target）增強雜交效率的效應。預先雜交之寡核苷酸片段和標的物DNA設計成與同軸堆疊之探針（Probe）序列完全互補。換而言之，先將一股DNA 寡核苷酸片段與探針上完全互補序列雜交後，再於同一探針序列上未雜交之部位，雜交另一股完全互補的DNA標的物。

預先雜交之寡核苷酸片段和DNA標的物，分別標定上明顯不同的螢光染色分子。藉由偵測螢光染色標定的訊號來研究，DNA堆疊和單獨雜交結構對於雜交效率的共同影響。結果顯示，預先雜交之寡核苷酸片段會增強於同一探針上後雜交DNA標的物的雜交效率。

關鍵字

核?酸 ； 標的物 ； 立體空間 ； 方位性 ； 雜交反應 ； 效率 ； 微陣列 ； 探針

並列摘要

The enhancement of hybridization efficiency of deoxyribose nucleic acid (DNA) targets by oigonucleotide pre-hybridization is studied on two sequence-inversed microarrayed probes. The sequences of both pre-hybridizing oligo and target DNA are designed to be fully complementary to their shared DNA probe in a coaxial stacking configuration; i.e. they hybridize immediately next to each other along the continuous complement probe strand. The pre-hybridizing oligo and target DNA are differentiated by labeling with two distinct fluorescent dyes, and the cooperative effect on hybridization efficiency is investigated through the comparison between the stacking and individual hybridization configurations based on the detection signals of the labeling dyes.
The results show that the pre-hybridization of a DNA oligo enhances the subsequent hybridization efficiency of the target DNA coupling onto the same probe.

並列關鍵字

Efficiency ； Hybridization ； DNA ； Orientated ； Target ； Microarray ； Steric ； Probe

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