He. Alcock et al., A simple method for PCR based analyses of immunohistochemically stained, microdissected, formalin fixed, paraffin wax embedded material, J CL PATH-M, 52(3), 1999, pp. 160-163
Citations number
9
Categorie Soggetti
Research/Laboratory Medicine & Medical Tecnology","Medical Research Diagnosis & Treatment
Microdissection was performed on sections cut from formalin fixed, paraffin
wax embedded archival material, which had been subjected to conventional i
mmunohistochemistry. Crude DNA extracts, which were obtained from these mic
rodissected samples by a simple microwave step, were then added directly to
amplification reactions. Analyses using a range of polymerase chain reacti
on (PCR) based techniques, including microsatellite repeat polymorphism ana
lysis at the NM23-H1 locus and sequencing of exons 5, 7, and 8 of the p53 g
ene, were performed successfully Universal PCR amplification was also carri
ed out on the microdissected material and probes suitable for use in compar
ative genomic hybridisation (CGH) were obtained in all cases. This techniqu
e will enable a range of effective genetic analyses to be carried out on sp
ecific subsets of cells that have been characterised previously by immunohi
stochemistry.