A NOVEL, RAPID METHOD FOR THE ISOLATION OF TERMINAL SEQUENCES FROM YEAST ARTIFICIAL CHROMOSOME (YAC) CLONES

被引：690
作者：
RILEY, J
BUTLER, R
OGILVIE, D
FINNIEAR, R
JENNER, D
POWELL, S
ANAND, R
SMITH, JC
MARKHAM, AF
机构：
[1] ICI Pharmaceuticals, Biotechnology Department, Cheshire SK10 4TG, Mereside, Alderley park, Macclesfield
来源：
NUCLEIC ACIDS RESEARCH | 1990年 / 18卷 / 10期
关键词：
D O I：
10.1093/nar/18.10.2887
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
The recent development of yeast artificial chromosome (YAC) vectors has provided a system for cloning fragments that are over ten times larger than those that can be cloned in more established systems. We have developed a method for the rapid isolation of terminal sequences from YAC clones. The YAC clone is digested with a range of restriction enzymes, a common linker is ligated to the DNA fragments and terminal sequences are amplified using a vector specific primer and a linker specific primer. Sequence data derived from these terminal specific products can be used to design primers for a further round of screening to isolate overlapping clones. The method also provides a convenient method of generating Sequence Tagged Sites for the mapping of complex genomes. © 1990 Oxford University Press.
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页码：2887 / 2890
页数：4
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