Microdissection as a means to verify allelic imbalance in tumour biology samples

Allelotypes (TP53, AFM051xd10 and alu-i1) in normal DNA and in DNA from paraffin-embedded tumours of a patient with a p53 germ-line mutation were compared in order to demonstrate LOH. Microdissection was applied in order to overcome difficulties with the interpretation of LOH data from a pelvic recurrence of a primary malignant histiocytoma. Furthermore, a rapid and simple boiling method was developed in order to reduce the loss of DNA usually occurring during traditional methods for DNA extraction. The conclusion drawn is that it is of utmost importance to use highly enriched fractions of tumour cells when performing LOH-studies. It is also shown that a rapid and simple boiling procedure is sufficient to release enough DNA of microdissection-enriched tumour cells for microsatellite analysis by PCR to detect allelic imbalance.