A SIMPLE AND EFFICIENT METHOD FOR MICRODISSECTION AND MICROFISH

A simple and efficient method for the dissection of (marker) chromosom es, (micro)nuclei, and chromosome regions is presented. Before microdi ssection, metaphases are overlaid with milli-Q water to rehydrate the chromosomes, which makes them soft and sticky. The dissected chromosom e fragments are dissolved without proteinase-K or topoisomerase treatm ent and directly amplified using a degenerate oligonucleotide primed p olymerase chain reaction (DOP-PCR). The advantages of this microFISH m ethod over previously reported methods are: (1) microdissection in thi s way is very fast; (2) a chromosome, marker, (micro)nucleus, or chrom osome region is collected as a whole using only one microneedle; (3) t he 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.