利用微陣列基因探針進行預雜交以增強DNA雜交效率

Efficiency Enhancement of DNA Hybridization by Pre-Hybridization on Microarrayed Gene Probes

姜凱仁(Kai-Ren Jiang)

指導教授 ： 吳瑞璋

中原大學/工學院/化學工程研究所/碩士(2009年)

https://doi.org/10.6840/cycu200901049

摘要

本研究利用兩種形態微陣列探針（Probe）與多樣的探針濃度，進行去氧核醣核苷酸（DNA）標的物（Target）預雜交反應（Pre-hybridization），以探討第二標的物樣品的雜交效率。使用三組經微陣列點樣之DNA探針和與其自身序列互補的標的物，每組包含一對DNA探針和一對與其自身序列互補並標定兩種易區分的螢光染色分子，經過特別設計的每個探針序列使標的物能分別互補於探針兩相鄰區域，分子空間效應對於雜交效率也透過比較單一及兩個標的物之間螢光訊號強度來探討。結果顯示在低探針濃度時有較好的雜交效率，且利用短序列DNA標的物進行第一次雜交反應可改善第二標的物鍵結至相同探針的雜交效率。然而第一標的物也發現因為第二標的物的鍵結而犧牲其自身訊號強度。

關鍵字

DNA微陣列 ； 分子立體障礙 ； 雜交效率 ； 基因晶片 ； DNA預雜交

並列摘要

The effect of pre-hybridizing genetic DNA target on the hybridization efficiency of the second target samples is studied on two probe configurations and various probe concentrations. Three sets of microarrayed DNA probes and their complementary samples are utilized. Each set contains a pair of DNA probes and a pair of their complementary samples labeled with two distinct fluorescent dyes. The sequence of each probe is especially designed so that two targets are simultaneously complementary to two adjacent sections of the probe. The molecular steric effect on the hybridization efficiency is investigated by comparing the dye signals between configurations of one-target and two-target hybridization scenarios. The results show that a low probe concentration gives better hybridization efficiency and the first-hybridization conducted by a shorter-size DNA target improves the hybridization efficiency of the second target coupling onto the same probe. The first DNA target is also found to sacrifice signal intensity for the second one.

並列關鍵字

hybridization efficiency ； DNA microarray ； molecular steric hindrance ； DNA pre-hybridization ； gene chips

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