A NEW METHOD FOR CONSTRUCTING NOTI LINKING AND BOUNDARY LIBRARIES USING A RESTRICTION TRAPPER

被引：17

作者：

HAYASHIZAKI, Y ^{[1
]}

HIROTSUNE, S ^{[1
]}

HATADA, I ^{[1
]}

TAMATSUKURI, S ^{[1
]}

MIYAMOTO, C ^{[1
]}

FURUICHI, Y ^{[1
]}

MUKAI, T ^{[1
]}

机构：

[1] NIPPON ROCHE RES CTR, DEPT MOLEC GENET, KAMAKURA, KANAGAWA 247, JAPAN

来源：

GENOMICS | 1992年 / 14卷 / 03期

关键词：

D O I：

10.1016/S0888-7543(05)80176-4

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

We have developed a novel method for constructing NotI linking and boundary libraries using a modified "solid-supported ligation primer" (restriction trapper). The restriction trapper could be used to purify the DNA fragments with a specific restriction enzyme cutting site(s) at their ends. The method uses a ligation and recutting reaction with double-stranded DNA ends of a hairpin-shaped oligolinker which is connected covalently to the surface of the latex beads. Selectivity is based on the specificity of the restriction enzyme for its recognition site, resulting in efficient purification. We applied this technique to the construction of high-quality NotI linking and NotI boundary libraries, which contain almost all the NotI sites of the genome and, in addition, are free of illegitimately ligated clones. © 1992 Academic Press, Inc. All rights reserved.

引用

页码：733 / 739

页数：7

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