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                                                                                            核酸適配體(Aptamer)與篩選文庫的合成

核酸適配體(Aptamer)是一類新型識別分子,其作用本質是一段單鏈DNA(脫氧核糖核酸)或者RNA(核糖核酸)分子折疊形成特定三維結構而  與生物靶標高親和力和高特異性結合。由于其分子量較小,可化學合成、穩定性好、沒有毒性等優點,在許多方面體現了傳統的免疫學與化學分子識別無可比擬的優  勢,在基礎研究及應用研究領域均呈現了快速發展的趨勢。

核酸適配體利用體外篩選技術-指數富集配體系統進化技術(Systematic evolution of ligands by   exponential enrichment, SELEX),從合成的核酸分子文庫中篩選得到的寡核苷酸片段。指數富集配體系統進化技術   (Selective expansion of ligands by exponential   enrichment,SELEX),指模擬自然進化人工篩選技術,首先體外化學合成一個隨機堿基數為n的單鏈寡核苷酸文庫,該文庫則含4n個不同的寡核苷酸序列,常用的寡核苷酸隨機序列含20-40個堿基,庫容量高達420-440(1012-24);隨機序列的兩端為隨后PCR循環時結合引物所必需的固定序列,由于這種隨機序列,而決定了庫中每條鏈自然形成的空間構象,即二級結構的多樣性,決定了庫中潛在地存在能與各種蛋白和低分子靶物質有親和力的核酸配體。一般篩選文庫的容量巨大(可達1015左右),理論上應用SELEX技術能篩選到自然界幾乎所有靶分子的適配子。

高靈敏、高通量的核酸適配體分子探針檢測新技術,為探尋發現惡性腫瘤的分子標志物新方法,建立有效的腫瘤早期預警、早期診斷與靶向監測技術平臺奠定基礎。

權陽生物目前提供如下產品和技術服務:

1.核酸適配體合成(參見常規引物合成、超長引物合成和高通量引物合成)

2.修飾核酸適配體合成(參見修飾引物合成與熒光探針引物合成)

3.隨機文庫產品

DNA Template (10 nmol)

5'-TAGGGAAGAGAAGGACATATGAT(N20, N30 or N40)TTGACTAGTACATGACCACTTGA-3'

N代表隨機合成A/T/C/G

正向擴增引物(20 nmol)

5'-TAGGGAAGAGAAGGACATATGAT-3'

反向擴增引物(20 nmol)

5'-TCAAGTGGTCATGTACTAGTCAA-3'

5' 生物素標記正向擴增引物(20 nmol)

  5' -Biotin-TAGGGAAGAGAAGGACATATGAT-3'

  5'生物素標記反向擴增引物(20 nmol)

  5' -Biotin-TCAAGTGGTCATGTACTAGTCAA-3'

T7啟動子引物(20 nmol):用于生成RNA篩選文庫

  5'-TTCAGGTAATACGACTCACTATAGGGAAGAGAAGGACATATGAT-3'

4.定制化的隨機文庫合成

引物服務和訂購:

根據您的需要請選擇相應的合成服務

Email訂購:請填寫引物合成訂購表(下載附件),聯系地址:[email protected]

發貨形式: 凍干粉或液體交貨

發貨COA 文件包括產品:
                                              1).引物序列;
                                              2). OD值;
                                              3).Tm 等詳細信息數據

 


【參考文獻】 


Wang RE, Wu H, Niu Y, Cai J. Improving the stability of aptamers by chemical modification. Curr Med Chem. 2011;18(27):4126-38.  


Hall B, Arshad S, Seo K, Bowman C, Corley M, Jhaveri SD,   Ellington AD. In vitro selection of RNA aptamers to a protein target by   filter immobilization. Curr Protoc Nucleic Acid Chem. 2010 Mar; Chapter   9:Unit 9.3.1-27.  


Kuwahara M, Sugimoto N. Molecular evolution of   functional nucleic acids with chemical modifications. Molecules. 2010   Aug 9;15(8):5423-44.  


Codrea V, Hayner M, Hall B, Jhaveri S, Ellington A. In   vitro selection of RNA aptamers to a small molecule target. Curr Protoc   Nucleic Acid Chem. 2010 Mar; Chapter 9:Unit 9.5.1-23.  


Mayer G, Höver T. In vitro selection of ssDNA aptamers using biotinylated target proteins. Methods Mol Biol. 2009;535:19-32.  


Gopinath SC. Methods developed for SELEX. Anal Bioanal Chem. 2007 Jan;387(1):171-82.  


Dausse E, Cazenave C, Rayner B, Toulmé JJ. In vitro   selection procedures for identifying DNA and RNA aptamers targeted to   nucleic acids and proteins. Methods Mol Biol. 2005;288:391-410.  


Nieuwlandt D. In vitro selection of functional nucleic acid sequences. Curr Issues Mol Biol. 2000 Jan;2(1):9-16.  


 


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