A simple and efficient method for microdissection and microFISH. Issue 4 (April 1998)

• Record Type: Journal Article
• Authors: Engelen, J J; Albrechts, J C; Hamers, G J; Geraedts, J P
• Is Part Of: Journal of medical genetics. Volume 35:Issue 4(1998)
• Page Start: 265
• Page End: 268
• Publication Date: 1998-04
• Subjects: Medical genetics -- Periodicals; 616.042
• DOI: 10.1136/jmg.35.4.265
• Languages: English
• ISSNs: 1468-6244

Abstract

Abstract : 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.