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PubMed · 9598716

A simple and efficient method for microdissection and microFISH.

J J Engelen·J C Albrechts·G J Hamers·J P Geraedts
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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.

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HumansIn Situ Hybridization, FluorescencePolymerase Chain ReactionJournal Article

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J J Engelen, J C Albrechts, G J Hamers, J P Geraedts. 1998. A simple and efficient method for microdissection and microFISH.. https://doi.org/10.1136/jmg.35.4.265

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