本實驗目的在測試微陣列系統的穩定性，如微陣列各點大小的均勻性如何，從組織或細胞獲得RNA的效率以及其後配對接合反應的效果。因為微陣列系統是新的技術，其中有些部分尚未徹底研究清楚，我們必須先行對系統本身仔細研究，了解其限制所在，如此才能對龐大基因資料作合理分析，得到最正確基因表現訊息。
微陣列均勻度測試實驗旨在瞭解點印過程均勻度，針與針之間的差異，以及所有針之間的差異，最後在資料分析時，我們能除去此差異性引進的實驗誤差。實驗設計為時間間隔下，每隔20分鐘點上40點，觀察探子(probe)的濃度變化，對均勻度有何影響。生物樣本RNA的製備，也是微陣列系統實驗重要的一環，必須有足夠量的RNA，方可進行螢光標定。實驗採用細胞株及胎盤(placenta)組織，所用細胞株為SKOV- 3，SKOV-3是一種人類卵巢癌(ovarian cancer)細胞。本實驗除了熟悉RNA的製備，也能比較不同生物樣本間，RNA含量的差異。另外也比較了採用total RNA以及mRNA進行配對接合反應時，兩者標幟上螢光分子的效率，期望能夠得到最佳的螢光偵測訊號。有了這些基礎實驗，我們才能對自己系統有一清楚認識，也因為是初探的實驗，微陣列系統之優化尚有許多值得研究與改進的空間。

The objective of this thesis is to study the stability of the cDNA mcroarray, such as the uniformity of each spot in the microarray, the efficiency of total RNA extraction from tissue or cells and the results of hybridization.Because cDNA microarray technology is new enough, we still have not known the details of it.So we must study the system carefully and find the limits.Then, reasonable analysis of huge gene information is to acquire the most correct gene expression patterns.
Knowing spots uniformity during printing, the statistic error of one pin and all variations between pins is the purpose of the microarray uniformity experiment.We can get rid of the systematic error in the final data analysis.The design of this experiment is to spot 40 spots every 20 minutes.Observation of the DNA probe concentration on the microarray have any effects on the system uniformity.The extraction of RNA form tissue or cells is also important in the microarray experiments.RNA used to be labeled fluorescent dye must have enough amount.We use cell line and placenta tissue to estimate the total RNA amount.SKOV3 cell line is one cell type of human ovarian caner.Besides knowing well the extraction

technology of total RNA, the difference of RNA amout in different samples can be compared.We also compare the results of hybridization by

total RNA or mRMA labeled fluorescent dye to see the intensity of spots is any different between two RNA types.

Because our microarray system is a new one, we can have much more knowledge of the system performance by these basic studies.This is a preliminary study and how to optimize the cDNA microarray technology is still to be worthy of another research and improvement.

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