寡核苷酸与固体支持物的共价连接。
Covalent attachment of oligonucleotides to solid supports.
作者信息
Ghosh S S, Musso G F
出版信息
Nucleic Acids Res. 1987 Jul 10;15(13):5353-72. doi: 10.1093/nar/15.13.5353.
Abstract
Coupling efficiencies for the covalent attachment of oligonucleotides (17-29 bases in length) to solid supports derivatized with alkyl-amino and -carboxylic functionalities have been determined. Attachment efficiencies of 60-80% were obtained for coated long-chain alkylamino controlled pore glass (CPG) supports. Similar efficiencies of immobilization were observed for carboxyl-bearing supports, which additionally exhibited lower levels of non-covalent binding. The extent of terminally linked oligonucleotide was determined to be 50-55% of the overall attachment in the carbodiimide-mediated coupling reaction of a 5'-aminohexyl phosphoramidate derivative of a 29-mer to Sephacryl carboxyl support. While lower overall efficiencies of attachment were obtained in the reaction with Sephacryl N-hydroxysuccinimide-activated carboxyl support, greater than 80% of this coupling results in end-attached oligonucleotides.
摘要
已测定了长度为17 - 29个碱基的寡核苷酸与用烷基氨基和羧基官能团衍生化的固体支持物共价连接的偶联效率。对于涂覆的长链烷基氨基可控孔径玻璃（CPG）支持物，获得了60 - 80%的连接效率。对于含羧基的支持物，观察到了类似的固定化效率，其还表现出较低水平的非共价结合。在将29聚体的5'-氨基己基亚磷酰胺衍生物与Sephacryl羧基支持物进行碳二亚胺介导的偶联反应中，末端连接的寡核苷酸的程度被确定为总连接量的50 - 55%。虽然在与Sephacryl N-羟基琥珀酰亚胺活化的羧基支持物的反应中获得的总连接效率较低，但这种偶联中超过80%的结果是末端连接的寡核苷酸。
相似文献
1
Covalent attachment of oligonucleotides to solid supports.寡核苷酸与固体支持物的共价连接。
Nucleic Acids Res. 1987 Jul 10;15(13):5353-72. doi: 10.1093/nar/15.13.5353.
2
Covalent attachment of hybridizable oligonucleotides to glass supports.可杂交寡核苷酸与玻璃载体的共价连接。
Anal Biochem. 1997 Apr 5;247(1):96-101. doi: 10.1006/abio.1997.2017.
3
Biotin-labeled oligonucleotides: enzymatic synthesis and use as hybridization probes.生物素标记的寡核苷酸：酶促合成及其作为杂交探针的应用
DNA. 1984 Jun;3(3):269-77. doi: 10.1089/dna.1.1984.3.269.
4
Phage T4 polynucleotide kinase.噬菌体T4多核苷酸激酶
Gene Amplif Anal. 1981;2:299-311.
5
Assessment of methods for covalent binding of nucleic acids to magnetic beads, Dynabeads, and the characteristics of the bound nucleic acids in hybridization reactions.核酸与磁珠、Dynabeads共价结合方法的评估以及杂交反应中结合核酸的特性。
Nucleic Acids Res. 1988 Nov 25;16(22):10861-80. doi: 10.1093/nar/16.22.10861.
6
End attachment of phenol-oligonucleotide conjugates to diazotized cellulose.苯酚 - 寡核苷酸缀合物与重氮化纤维素的末端连接。
Bioconjug Chem. 1993 Sep-Oct;4(5):380-5. doi: 10.1021/bc00023a013.
7
Immobilization of oligonucleotides onto a glass support via disulfide bonds: A method for preparation of DNA microarrays.通过二硫键将寡核苷酸固定在玻璃载体上：一种制备DNA微阵列的方法。
Anal Biochem. 1999 Jan 1;266(1):23-30. doi: 10.1006/abio.1998.2857.
8
Oligo-microarray based on oligonucleotide immobilization on glass surface modified with activated acrylic acid-co-acrylamide copolymer.基于寡核苷酸固定在经活性丙烯酸-丙烯酰胺共聚物修饰的玻璃表面的寡核苷酸微阵列。
J Biomed Mater Res B Appl Biomater. 2008 Oct;87(1):67-72. doi: 10.1002/jbm.b.31069.
9
Rapid esterification of nucleosides to solid-phase supports for oligonucleotide synthesis using uronium and phosphonium coupling reagents.使用鎓盐和鏻盐偶联试剂将核苷快速酯化到固相载体上用于寡核苷酸合成。
Bioconjug Chem. 1999 Nov-Dec;10(6):1051-7. doi: 10.1021/bc990063a.
10
An improved procedure for derivatization of controlled-pore glass beads for solid-phase oligonucleotide synthesis.一种用于固相寡核苷酸合成的可控孔玻璃珠衍生化的改进方法。
Nucleic Acids Res. 1990 Jul 11;18(13):3813-21. doi: 10.1093/nar/18.13.3813.
引用本文的文献
1
A gold nanoparticle-protein G electrochemical affinity biosensor for the detection of SARS-CoV-2 antibodies: a surface modification approach.一种基于金纳米颗粒-蛋白 G 电化学亲和生物传感器的 SARS-CoV-2 抗体检测方法：一种表面修饰方法。
Sci Rep. 2022 Jul 27;12(1):12850. doi: 10.1038/s41598-022-17219-7.
2
Scanometric Detection of Tomato Leaf Curl New Delhi Viral DNA Using Mono- and Bifunctional AuNP-Conjugated Oligonucleotide Probes.使用单功能和双功能金纳米颗粒偶联寡核苷酸探针扫描检测番茄卷叶新德里病毒DNA
ACS Omega. 2019 Jun 11;4(6):10094-10107. doi: 10.1021/acsomega.9b00340. eCollection 2019 Jun 30.
3
Drop drying on surfaces determines chemical reactivity - the specific case of immobilization of oligonucleotides on microarrays.表面上的滴干法决定化学反应活性——寡核苷酸固定在微阵列上的具体情况。
BMC Biophys. 2013 Jun 12;6:8. doi: 10.1186/2046-1682-6-8.
4
Fabrication of oligonucleotide and protein arrays on rigid and flexible substrates coated with reactive polymer multilayers.在涂覆有反应性聚合物多层膜的刚性和柔性基底上制造寡核苷酸和蛋白质阵列。
ACS Appl Mater Interfaces. 2013 Jan 23;5(2):351-9. doi: 10.1021/am302285n. Epub 2012 Dec 28.
5
Northern blot detection of endogenous small RNAs (approximately14 nt) in bacterial total RNA extracts.细菌总 RNA 提取物中内源性小 RNA（约 14nt）的 Northern blot 检测。
Nucleic Acids Res. 2010 Aug;38(14):e147. doi: 10.1093/nar/gkq437. Epub 2010 May 26.
6
Carbodiimide-mediated cross-linking of RNA to nylon membranes improves the detection of siRNA, miRNA and piRNA by northern blot.碳二亚胺介导的RNA与尼龙膜交联可提高Northern印迹法对小干扰RNA（siRNA）、微小RNA（miRNA）和Piwi相互作用RNA（piRNA）的检测效果。
Nucleic Acids Res. 2007;35(8):e60. doi: 10.1093/nar/gkm112. Epub 2007 Apr 2.
7
Solid phase DNA amplification: characterisation of primer attachment and amplification mechanisms.固相DNA扩增：引物连接与扩增机制的表征
Nucleic Acids Res. 2000 Oct 15;28(20):E87. doi: 10.1093/nar/28.20.e87.
8
Covalent attachment of synthetic DNA to self-assembled monolayer films.合成DNA与自组装单分子层膜的共价连接。
Nucleic Acids Res. 1996 Aug 1;24(15):3031-9. doi: 10.1093/nar/24.15.3031.
9
Real time kinetics of restriction endonuclease cleavage monitored by fluorescence resonance energy transfer.
Nucleic Acids Res. 1994 Aug 11;22(15):3155-9. doi: 10.1093/nar/22.15.3155.
10
Direct fluorescence analysis of genetic polymorphisms by hybridization with oligonucleotide arrays on glass supports.通过与玻璃载体上的寡核苷酸阵列杂交对基因多态性进行直接荧光分析。
Nucleic Acids Res. 1994 Dec 11;22(24):5456-65. doi: 10.1093/nar/22.24.5456.
本文引用的文献
1
Derivatization of unprotected polynucleotides.未保护的多核苷酸的衍生化
Nucleic Acids Res. 1983 Sep 24;11(18):6513-29. doi: 10.1093/nar/11.18.6513.
2
Catalysis of accurate poly(C)-directed synthesis of 3'-5'-linked oligoguanylates by Zn2+.锌离子催化以聚胞嘧啶为模板精确合成3'-5'连接的寡聚鸟苷酸。
J Mol Biol. 1980 Dec 25;144(4):567-77. doi: 10.1016/0022-2836(80)90337-x.
3
Hybridization of nucleic acids immobilized on solid supports.固定在固体支持物上的核酸杂交。
Anal Biochem. 1984 May 1;138(2):267-84. doi: 10.1016/0003-2697(84)90808-x.
4
Immobilization of denatured DNA to macroporous supports: I. Efficiency of different coupling procedures.变性DNA固定于大孔载体：I. 不同偶联方法的效率
Nucleic Acids Res. 1982 Nov 25;10(22):7163-80. doi: 10.1093/nar/10.22.7163.
5
[Synthesis of the C-terminal hexapeptide amide of secretin on a new solid support].[在一种新型固相载体上合成促胰液素C末端六肽酰胺]
Hoppe Seylers Z Physiol Chem. 1971 May;352(5):759-60.
6
Color test for detection of free terminal amino groups in the solid-phase synthesis of peptides.用于检测肽固相合成中游离末端氨基的颜色测试。
Anal Biochem. 1970 Apr;34(2):595-8. doi: 10.1016/0003-2697(70)90146-6.
7
The monitoring of reactions in solid-phase peptide synthesis with picric acid.用苦味酸监测固相肽合成中的反应。
Anal Chim Acta. 1972 Jan;58(1):248-9. doi: 10.1016/S0003-2670(00)86882-8.
8
Studies on polynucleotides. The linkage of deoxyribopolynucleotide templates to cellulose and its use in their replication.多核苷酸研究。脱氧核糖多核苷酸模板与纤维素的连接及其在复制中的应用。
J Biol Chem. 1974 Aug 25;249(16):5213-21.
9
Detection of specific DNA sequences with short biotin-labeled probes.用短生物素标记探针检测特定DNA序列。
DNA. 1985 Aug;4(4):327-31. doi: 10.1089/dna.1985.4.327.
10
Biotin-labeled synthetic oligodeoxyribonucleotides: chemical synthesis and uses as hybridization probes.生物素标记的合成寡脱氧核糖核苷酸：化学合成及其作为杂交探针的用途。
Nucleic Acids Res. 1985 Mar 11;13(5):1529-41. doi: 10.1093/nar/13.5.1529.
Zotero 插件