Differential polymerase chain reaction: a technical comparison of three methods for the detection of CDK4 gene amplification in glioblastomas.
Among the different techniques used to detect oncogene amplification in tumor DNA, Southern blot and differential PCR have been the most frequently used. We report on a technical comparison of three different methods to detect gene amplification by differential PCR: ethidium bromide staining, silver staining (both after standard differential PCR), and fluorescent differential PCR. We explored the relative densitometric measure of a 119 bp fragment of the CDK4 gene versus an 82 bp fragment of the IFNG gene. In total agreement with previous studies carried out by Southern blot and differential PCR by other authors, we were able to detect CDK4 amplification in 3 of the 21 glioblastomas (14%), but only by the fluorescent differential PCR method. In conclusion, fluorescent differential PCR is more sensitive than standard differential PCR for detection oncogene amplification in tumor DNAs.
Department of Genetics, University of Navarre, Pamplona, Spain.
X
X
Fan
L
L
Gómez
M
M
Nistal
L
L
Sierrasesúmaga
J S
JS
Castresana
D001932:Brain Neoplasms / Q000235:genetics* / Q000378:metabolism; D051358:Cyclin-Dependent Kinase 4; D018844:Cyclin-Dependent Kinases / Q000032:analysis / Q000235:genetics*; D004273:DNA, Neoplasm / Q000032:analysis; D005069:Evaluation Studies as Topic; D005784:Gene Amplification; D005909:Glioblastoma / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis; D007962:Leukocytes / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D011518:Proto-Oncogene Proteins
D004273:DNA, Neoplasm; D011518:Proto-Oncogene Proteins; D007371:Interferon-gamma; C495900:CDK4 protein, human; D051358:Cyclin-Dependent Kinase 4; D018844:Cyclin-Dependent Kinases