[Detection of point mutations on the Ki-ras gene using the PCR technique].
Present study was undertaken to detect Ki-ras point mutation at codon 12 in pancreatic adenocarcinomas (CaP) using the polymerase chain reaction-restriction fragment lengths polymorphism (PCR-RFLP). Three modifications of PCR-RFLP were performed with a mismatched primers creating a recognition site with only one allelic from (wild or mutated). Using two-step PCR-RFLP and two modifications of one-step PCR-RFLP we examined 5 resected adenocarcinomas of pancreas, 6 pancreatic juices and one DNA sample from peripheral blood of patient with generalized stadium of CaP. We compare all techniques and conclude, that the very sensitive two step PCR-RFLP is a suitable method for detection point mutations and eliminates the need for either oligonucleotide hybridization or DNA sequencing.
Janda
J
J
I. interní klinika 1. LF UK a VFN, Praha.
Krechler
T
T
Dufek
V
V
Klener
P
P
D000230:Adenocarcinoma / Q000235:genetics*; D011905:Genes, ras / Q000235:genetics*; D006801:Humans; D010190:Pancreatic Neoplasms / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length