隨著阿拉伯芥被完整解序後，其他植物基因組的定序工程也陸續完成，例如：水稻、玉米、馬鈴薯等。2012年，在多國學者的合作下更完成了蕃茄基因組定序 (真染色質部分) 工作，這項成果更帶動了涵蓋許多經濟作物的茄科植物的研究與發展。若依照傳統定序的方式，必須先建立物種基因庫和遺傳圖譜才能將物種序列進行排序，但基因庫和遺傳圖譜的建構十分耗時。因此若想發展一項新技術，在具備染色體核型圖譜 (karyotyping) 的前提下，單純以顯微切割技術再結合phi29 DNA聚合酶特殊多重置換的機制，使用illustraTM GenomiPhi Amplification kit (GE Healthcare) 達到大量擴增，再將擴增產物進行解序，可省去基因庫與遺傳圖譜建構的時間。為了測定此技術可行性，將顯微切割擴增後的產物 (6L) 運用螢光原位雜交技術 (Fluorescence in situ hybridization) 回標至蕃茄第六條染色體上。實驗結果發現，6L與重複性序列45S rDNA訊號位置重疊，表示6L並非本實驗預期中第六條染色體長臂片段，也意味著對於目標區域進行顯微切割的操作過程中，準確度及穩定性具有決定性因素。因此透過本研究初探，此項技術還有許多改進之處，例如：顯微切割方式、顯微切割用染色體製備和切割後DNA品質的確認等。希望將來可以建立更完整的原位顯微切割技術，解決並改善傳統定序耗時的問題，增進未來對基因組定序的便利性與精確性。

Genomiphi amplification is one of the whole genome amplification (WGA) tools. It work on generating microgram quantities from low copy number DNA or degraded DNA with multiple displacement amplification (MDA) mechanisms of phi29 DNA polymerase. We described an innovation that using microdissection technique combined illustraTM GenomiPhi Amplification kit (GE Healthcare) to solve the obstacles of conventional sequencing in spending time to construct gene library and genetic map. In order to evaluate the feasibility of this strategy, we chose long arm region of tomato chromosome 6 as material for needle-microdissection and reconfirmed WGA-DNA (dissected segment, 6L) by fluorescence in situ hybridization (FISH). According to the results of 6L and 45S rDNA show the same loci, which means that is not expected results. Therefore, the procedure of this strategy needs to be re-examined. We hope improved technique can get more accuracy and convenient for genome sequencing.

李思元和莊以光，DNA定序技術之演進與發展 (2010)

薛豪彥，蘭嶼姬蝴蝶蘭粗絲期染色體螢光原位雜交與核型分析，國立臺灣大學農藝研究所碩士論文 (2012)

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