A simple and efficient method for microdissection and microFISH

被引：42

作者：

Engelen, JJM ^{[1
]}

Albrechts, JCM ^{[1
]}

Hamers, GJH ^{[1
]}

Geraedts, JPM ^{[1
]}

机构：

[1] Univ Limburg, Dept Mol Genet & Cell Biol, NL-6200 MD Maastricht, Netherlands

来源：

JOURNAL OF MEDICAL GENETICS | 1998年 / 35卷 / 04期

关键词：

microdissection; DOP-PCR; fluorescence in situ hybridisation;

D O I：

10.1136/jmg.35.4.265

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

A simple and efficient method for the dissection of (marker) chromosomes, (micro)nuclei, and chromosome regions is presented. Before microdissection, metaphases are overlaid with milli-Q water to rehydrate the chromosomes, which makes them soft and sticky. The dissected chromosome fragments are dissolved without proteinase-K or topoisomerase treatment and directly amplified using a degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR). The advantages of this microFISH method over previously reported methods are: (1) microdissection in this way is very fast; (2) a chromosome, marker, (micro)nucleus, or chromosome region is collected as a whole using only one microneedle; (3) the dissected material sticks tightly to the needle without the risk of getting lost; (4) no Sequenase is used in the DOP-PCR reaction which reduces the risk of contamination.

引用

页码：265 / 268

页数：4

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