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                                                                                            核酸适配体(Aptamer)与筛选文库的合成

核酸适配体(Aptamer)是一类新型识别分子,其作用本质是一段单链DNA(脱氧核糖核酸)或者RNA(核糖核酸)分子折叠形成特定三维结构而  与生物靶标高亲和力和高特异性结合。由于其分子量较小,可化学合成、稳定性好、没有毒性等优点,在许多方面体现了传统的免疫学与化学分子识别无可比拟的优  势,在基础研究及应用研究领域均呈现了快速发展的趋势。

核酸适配体利用体外筛选技术-指数富集配体系统进化技术(Systematic evolution of ligands by   exponential enrichment, SELEX),从合成的核酸分子文库中筛选得到的寡核苷酸片段。指数富集配体系统进化技术   (Selective expansion of ligands by exponential   enrichment,SELEX),指模拟自然进化人工筛选技术,首先体外化学合成一个随机碱基数为n的单链寡核苷酸文库,该文库则含4n个不同的寡核苷酸序列,常用的寡核苷酸随机序列含20-40个碱基,库容量高达420-440(1012-24);随机序列的两端为随后PCR循环时结合引物所必需的固定序列,由于这种随机序列,而决定了库中每条链自然形成的空间构象,即二级结构的多样性,决定了库中潜在地存在能与各种蛋白和低分子靶物质有亲和力的核酸配体。一般筛选文库的容量巨大(可达1015左右),理论上应用SELEX技术能筛选到自然界几乎所有靶分子的适配子。

高灵敏、高通量的核酸适配体分子探针检测新技术,为探寻发现恶性肿瘤的分子标志物新方法,建立有效的肿瘤早期预警、早期诊断与靶向监测技术平台奠定基础。

权阳生物目前提供如下产品和技术服务:

1.核酸适配体合成(参见常规引物合成、超长引物合成和高通量引物合成)

2.修饰核酸适配体合成(参见修饰引物合成与荧光探针引物合成)

3.随机文库产品

DNA Template (10 nmol)

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

N代表随机合成A/T/C/G

正向扩增引物(20 nmol)

5'-TAGGGAAGAGAAGGACATATGAT-3'

反向扩增引物(20 nmol)

5'-TCAAGTGGTCATGTACTAGTCAA-3'

5' 生物素标记正向扩增引物(20 nmol)

  5' -Biotin-TAGGGAAGAGAAGGACATATGAT-3'

  5'生物素标记反向扩增引物(20 nmol)

  5' -Biotin-TCAAGTGGTCATGTACTAGTCAA-3'

T7启动子引物(20 nmol):用于生成RNA筛选文库

  5'-TTCAGGTAATACGACTCACTATAGGGAAGAGAAGGACATATGAT-3'

4.定制化的随机文库合成

引物服务和订购:

根据您的需要请选择相应的合成服务

Email订购:请填写引物合成订购表(下载附件),联系地址:[email protected]

发货形式: 冻干粉或液体交货

发货COA 文件包括产品:
                                              1).引物序列;
                                              2). OD值;
                                              3).Tm 等详细信息数据

 


【参考文献】 


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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