Method for detecting mutations of H-ras in the presence of an excess of normal cells

摘要

Method which consists in the detection of mutations of the oncogene H-ras by means of the amplification of a fragment of 308 base pairs which delimit codon 12, which is most frequently mutated, digestion with a restriction enzyme which selectively suppresses the non-mutated chains, leaving those in which the restriction site is not present due to mutation in this codon, and a novel amplification of a fragment of 129 base pairs. Because the enzyme does not suppress 100% of the non-mutated chains, the product is again digested with the same enzyme and is blotted on a gel. The appearance of a band of 129 base pairs (resistant to suppression with the restriction enzyme) is interpreted as the presence of a mutation of codon 12 of the H-ras gene. IMAGE