DNA and RNA isolated from tissues processed by microwave-accelerated apparatus MFX-800-3 are suitable for subsequent PCR and Q-RT-PCR amplification.
Over the past decade, methods of molecular biology have appeared in diagnostic pathology and are routinely applied on formalin-fixed, paraffin-embedded histological samples, processed via conventional embedding methods. Due to its reagent- and cost-effectiveness, embedding techniques that utilize microwave acceleration in one or more steps of histoprocessing are increasingly used by numerous laboratories. The demand arises that tissues processed this way should also be suitable for the requirements of molecular pathology. In this study, both conventionally embedded and MFX-800-3 machine-processed tissue samples from the same source were used for isolation of DNA and RNA and for performing PCR and real-time PCR. PCR amplification of the beta-globin gene, as well as the real-time PCR amplification of the ABL mRNA was successful in all cases. Our conclusion is that samples processed by the vacuum assisted automatic microwave histoprocessor MFX-800-3 are perfectly applicable for DNA and RNA isolation and provide appropriate templates for further PCR and realtime PCR studies.
Bödör
Csaba
C
1st Department of Pathology and Experimental Cancer Research, Faculty of Medicine, Semmelweis University, Budapest, H-1085, Hungary. bodor@korb1.sote.hu
Schmidt
Otto
O
Csernus
Balázs
B
Rajnai
Hajnalka
H
Szende
Béla
B
D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000302:isolation & purification* / Q000378:metabolism; D004273:DNA, Neoplasm / Q000302:isolation & purification / Q000378:metabolism; D005704:Gallbladder / Q000378:metabolism; D005914:Globins / Q000235:genetics; D006801:Humans; D008099:Liver / Q000378:metabolism; D008872:Microwaves; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012313:RNA / Q000302:isolation & purification* / Q000378:metabolism; D012334:RNA, Neoplasm / Q000302:isolation & purification / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D016610:Tissue Embedding / Q000295:instrumentation*; D014618:Vacuum
D004273:DNA, Neoplasm; D012334:RNA, Neoplasm; D012313:RNA; D005914:Globins; D004247:DNA