Rapid identification of yeast artificial chromosome clones by matrix pooling and crude Iysate PCR
نویسندگان
چکیده
منابع مشابه
USE OF VECTORETTE AND SUBVECTORETTE PCR FOR THE ISOLATION OF TERMINAL SEQUENCES FROM Y EAST ARTIFICIAL CHROMOSOME (YAC) CLONES
Development of yeast artificial chromosome (Y AC) vectors, molecular cloning of large segments of chromosomal DNA, and their propagation in yeast cells has become feasible. Overlapping Y AC provides a route to the development of physical maps of entire mammalian chromosomes. A rapid method was developed to isolate and sequence termini of Y AC inserts quickly. The Y AC clone is digested wit...
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The cloning of large DNA fragments of hundreds of kilobases in Yeast artificial chromosomes, has simplified the analysis of regions of the genome previously cloned by cosmid walking. The mapping of expressed sequences within cosmid contigs has relied on the association of genes with sequence motifs defined by rare-cutting endonucleases, and the identification of sequence conservation between sp...
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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 frag...
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In this article, we used PCR analysis with arbitrary primers (AP-PCR) to screen for overlapping bacterial artificial chromosome (BAC) clones and assembly of contigs. A rice BAC library with three genome equivalents was used to prepare pooled BAC DNA. Twenty-two arbitrary primers were used to survey the pooled BAC DNAs and individual BAC DNAs. Each primer identified 1-10 loci, and the average wa...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1990
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/18.23.7191