RT Journal Article
SR Electronic
T1 A simple and efficient method for microdissection and microFISH.
JF Journal of Medical Genetics
JO J Med Genet
FD BMJ Publishing Group Ltd
SP 265
OP 268
DO 10.1136/jmg.35.4.265
VO 35
IS 4
A1 Engelen, J J
A1 Albrechts, J C
A1 Hamers, G J
A1 Geraedts, J P
YR 1998
UL http://jmg.bmj.com/content/35/4/265.abstract
AB 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.