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通过将溴化氰活化的纤维素与5'-氨基末端寡核苷酸和聚（pdT）偶联来制备与固相支持物相连的聚脱氧核苷酸。

Preparation of polydeoxynucleotides linked to a solid support by coupling CNBr-activated cellulose with 5'-NH2-terminated oligo and poly(pdT)'s.

作者信息

Clerici L, Campagnari F, de Rooij J F, van Boom J H

出版信息

Nucleic Acids Res. 1979 Jan;6(1):247-58. doi: 10.1093/nar/6.1.247.

DOI:10.1093/nar/6.1.247
PMID:424291
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC327686/
Abstract

Synthetically-prepared 5'-NH2-dT(pdT)n oligomers (66,n=4 or 7) were immobilized on cyanogen bromide activated cellulose. The influence of temperature, pH, and ionic strength on the rate of the coupling process was studied. The oligomer 5'-NH2-DT(pdT)8 could be elongated enzymatically to the polymers 5'-NH2-dT(pdT)n (n=20, 51 and 84), which could be immobilized on cellulose. The cellulose-NH-dT(pdT)84 polymer thus obtained could be assembled to a new solid-state polymer e.g. poly(dA)290 . poly(/3H/dT)200, poly(dT)85-cellulose which, in turn, was a very convenient substrate for assaying DNA-ligase.

摘要

合成制备的5'-NH2-dT(pdT)n寡聚物（66，n = 4或7）被固定在溴化氰活化的纤维素上。研究了温度、pH值和离子强度对偶联过程速率的影响。寡聚物5'-NH2-DT(pdT)8可以通过酶促反应延长为聚合物5'-NH2-dT(pdT)n（n = 20、51和84），这些聚合物可以固定在纤维素上。由此得到的纤维素-NH-dT(pdT)84聚合物可以组装成一种新的固态聚合物，例如聚(dA)290。聚(/3H/dT)200，聚(dT)85-纤维素，反过来，它是用于检测DNA连接酶的非常方便的底物。

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Preparation of polydeoxynucleotides linked to a solid support by coupling CNBr-activated cellulose with 5'-NH2-terminated oligo and poly(pdT)'s.通过将溴化氰活化的纤维素与5'-氨基末端寡核苷酸和聚（pdT）偶联来制备与固相支持物相连的聚脱氧核苷酸。
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本文引用的文献

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Some general methods of preparing affinity columns.制备亲和柱的一些常规方法。
Nucleic Acids Res. 1974 Nov;1(11):1587-99. doi: 10.1093/nar/1.11.1587.
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Polynucleotide cellulose as a substrate for a polynucleotide ligase induced by phage T4.多核苷酸纤维素作为噬菌体T4诱导的多核苷酸连接酶的底物。
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Covalent attachment of nucleic acids to agarose for affinity chromatography.用于亲和色谱的核酸与琼脂糖的共价连接。
Biochemistry. 1971 Feb 2;10(3):424-7. doi: 10.1021/bi00779a011.
7
Deoxynucleotide-polymerizing enzymes of calf thymus gland. V. Homogeneous terminal deoxynucleotidyl transferase.小牛胸腺的脱氧核苷酸聚合酶。V. 均一的末端脱氧核苷酸转移酶。
J Biol Chem. 1971 Feb 25;246(4):909-16.
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Determination of molecular weight distributions of DNA by means of sedimentation in a sucrose gradient.通过蔗糖梯度沉降法测定DNA的分子量分布
Anal Biochem. 1969 Oct 15;32(1):14-30. doi: 10.1016/0003-2697(69)90100-6.
9
Oligodeoxythymidylate: polydeoxyadenylate and oligodeoxyadenylate: polydeoxythymidylate interactions.寡聚脱氧胸苷酸：聚脱氧腺苷酸以及寡聚脱氧腺苷酸：聚脱氧胸苷酸相互作用
Biochemistry. 1969 Oct;8(10):3928-36. doi: 10.1021/bi00838a008.
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Purification of a DNA polymerase from calf thymus nuclei.从小牛胸腺细胞核中纯化一种DNA聚合酶。
Biochim Biophys Acta. 1972 Jul 31;272(4):526-38. doi: 10.1016/0005-2787(72)90508-4.