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将非放射性标记物连接到合成寡脱氧核糖核苷酸5'末端的高效方法。

Efficient methods for attaching non-radioactive labels to the 5' ends of synthetic oligodeoxyribonucleotides.

作者信息

Agrawal S, Christodoulou C, Gait M J

出版信息

Nucleic Acids Res. 1986 Aug 11;14(15):6227-45. doi: 10.1093/nar/14.15.6227.

DOI:10.1093/nar/14.15.6227
PMID:3748808
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC311633/
Abstract

The syntheses are described of two types of linker molecule useful for the specific attachment of non-radioactive labels such as biotin and fluorophores to the 5' terminus of synthetic oligodeoxyribonucleotides. The linkers are designed such that they can be coupled to the oligonucleotide as a final step in solid-phase synthesis using commercial DNA synthesis machines. Increased sensitivity of biotin detection was possible using an anti-biotin hybridoma/peroxidase detection system.

摘要

描述了两种连接分子的合成方法，这两种连接分子可用于将生物素和荧光团等非放射性标记物特异性连接到合成寡脱氧核糖核苷酸的5'末端。这些连接分子的设计使得它们能够在使用商用DNA合成仪进行固相合成的最后一步与寡核苷酸偶联。使用抗生物素杂交瘤/过氧化物酶检测系统可以提高生物素检测的灵敏度。

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Nucleic Acids Res. 1986 Aug 11;14(15):6227-45. doi: 10.1093/nar/14.15.6227.
2
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本文引用的文献

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PARTIAL PURIFICATION OF SOLUBLE RNA.可溶性核糖核酸的部分纯化
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New fluorescent hydrazide reagents for the oxidized 3'-terminus of RNA.用于RNA氧化3'-末端的新型荧光酰肼试剂。
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Rapid synthesis of oligodeoxyribonucleotides VI. Efficient, mechanised synthesis of heptadecadeoxyribonucleotides by an improved solid phase phosphotriester route.寡脱氧核糖核苷酸的快速合成VI。通过改进的固相磷三酯路线高效、机械化合成十七聚脱氧核糖核苷酸。
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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
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In situ activation of bis-dialkylaminophosphines--a new method for synthesizing deoxyoligonucleotides on polymer supports.双二烷基氨基膦的原位活化——一种在聚合物载体上合成脱氧寡核苷酸的新方法。
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6
Use of ribonucleosides as protecting groups in synthesis of polynucleotides with phosphorylated terminals.核糖核苷在合成具有磷酸化末端的多核苷酸中作为保护基团的应用。
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7
Polymer support oligonucleotide synthesis XVIII: use of beta-cyanoethyl-N,N-dialkylamino-/N-morpholino phosphoramidite of deoxynucleosides for the synthesis of DNA fragments simplifying deprotection and isolation of the final product.聚合物载体寡核苷酸合成 XVIII：脱氧核苷的β-氰基乙基-N，N-二烷基氨基-/N-吗啉代亚磷酰胺用于 DNA 片段合成，简化最终产物的脱保护和分离
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Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes.生物素标记多核苷酸的酶促合成：新型核酸亲和探针。
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10
Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots.用于可视化与固定在硝酸纤维素膜上的DNA或RNA杂交的生物素标记DNA探针的快速灵敏比色法：生物印迹法。
Proc Natl Acad Sci U S A. 1983 Jul;80(13):4045-9. doi: 10.1073/pnas.80.13.4045.