A reliable PCR amplification method for microdissected tumor cells obtained from paraffin-embedded tissue.
We report a reliable method for PCR (polymerase chain reaction) amplification of genomic DNA from PET. This method uses DNA extraction with the QIAquick kit and amplification with AmpliTaq Gold. Amplification of up to 959 bp from PET was achieved with this combination which exceeds the current reported upper limit of 800 bp. In summary, the gradual activation of the AmpliTaq Gold during thermal cycling allows both for higher-fidelity and higher-throughput PCR amplification from PET. The use of the QIAquick kit for DNA purification of PET is sensitive, reproducible and suitable for management of a high number of samples.
Akalu
A
A
Institute for Genetic Medicine, USC School of Medicine, Los Angeles, CA 90033, USA.
Reichardt
J K
JK
D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase / Q000235:genetics; D004273:DNA, Neoplasm / Q000032:analysis*; D006801:Humans; D008297:Male; D009154:Mutation; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*; D011471:Prostatic Neoplasms / Q000737:chemistry / Q000235:genetics*; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D019914:Taq Polymerase / Q000737:chemistry
D004273:DNA, Neoplasm; D011933:Reagent Kits, Diagnostic; D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase; D019914:Taq Polymerase