CHROMOSOMAL INSITU HYBRIDIZATION USING YEAST ARTIFICIAL CHROMOSOMES

被引：29

作者：

SELLERI, L

HERMANSON, GG

EUBANKS, JH

EVANS, GA

机构：

[1] SALK INST BIOL STUDIES,MOLEC GENET LAB,POB 85800,LA JOLLA,CA 92037

[2] SALK INST BIOL STUDIES,CTR HUMAN GENOME RES,LA JOLLA,CA 92037

来源：

GENETIC ANALYSIS-BIOMOLECULAR ENGINEERING | 1991年 / 8卷 / 02期

基金：

美国国家卫生研究院;

关键词：

D O I：

10.1016/1050-3862(91)90050-2

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Large DNA fragment cloning methods using yeast artificial chromosomes (YACs) have vastly improved the strategies for constructing physical maps of regions of complex genomes, as well as for isolating and cloning genes important for human disease. We present here a simple and rapid method for carrying out in situ hybridization to metaphase chromosomes using isolated YAC clones by labeling DNA directly in agarose gel slices. Nonisotopic labeling and chromosomal in situ hybridization can be used to determine the chromosomal localization of individual YAC clones on human metaphase chromosomes. This method can also be used to characterize YAC clones consisting of single fragments from those that contain concatamerized, and thus artifactual, inserts. This technique also offers a valuable tool to study consistent translocations in neoplastic diseases by identifying YACs that span a specific chromosomal breakpoint.

引用

页码：59 / 66

页数：8

共 42 条

[1] CONSTRUCTION OF YEAST ARTIFICIAL CHROMOSOME LIBRARIES WITH LARGE INSERTS USING FRACTIONATION BY PULSED-FIELD GEL-ELECTROPHORESIS [J].