The application of microwave denaturation in comparative genomic hybridization

被引:12
作者
deMeulemeester, M [1 ]
Vink, A [1 ]
Jakobs, M [1 ]
Hermsen, M [1 ]
Steenman, M [1 ]
Slater, R [1 ]
Dietrich, A [1 ]
Mannens, M [1 ]
机构
[1] FREE UNIV AMSTERDAM HOSP,DEPT PATHOL,AMSTERDAM,NETHERLANDS
来源
GENETIC ANALYSIS-BIOMOLECULAR ENGINEERING | 1996年 / 13卷 / 05期
关键词
microwave; denaturation; comparative genomic hybridization; FISH;
D O I
10.1016/S1050-3862(96)00162-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Comparative genomic hybridization (CGH) is a powerful tool for analyzing unbalanced chromosomal rearrangements in a variety of tissues. However, the reproducibility of the technique is poor. We have developed an alternative protocol involving microwave denaturation of the metaphase chromosome preparations prior to the hybridization step. The advantage of this method for CGH is the retention of the morphology of the chromosomes and hence an improved chromosome banding pattern. Furthermore, it results in a consistently strong hybridization which is not dependent on the batch of lymphocytes used to obtain the metaphase chromosome spreads. This procedure has also proved to be applicable to nucleic acid hybridizations in general. The protocol. its application and the results of this method in CGH is discussed. Furthermore preliminary results of this method in paint and DNA probe hybridizations to chromosome spreads and to RNA in tissue sections are presented. Copyright (C) 1996 Elsevier Science B.V.
引用
收藏
页码:129 / 133
页数:5
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