Patent ID: 7892742

Claim:
A method for detecting a point mutation in a gene of interest comprising: a) performing a PCR reaction on an extract of DNA obtained from a plant containing the gene of interest using a pair of PCR primers which are capable of amplifying the region surrounding and encompassing the mutation; b) digesting the PCR product of step (a) with at least one restriction enzyme, which differentiates between the wild type sequence and the sequence containing the mutation, in order to obtain a restriction digest; c) analysing the digested PCR product of step (b) in order to identify those DNA extracts indicative of the presence of the mutation; characterised in that (i) the 3′ ends of the PCR primers are separated by 3 or fewer nucleotides, (ii) more than one mutation of interest is present in the 3 or fewer nucleotides between the 3′ ends of the primers and (iii) the product of the PCR reaction is such that each mutations gives a different restriction profile when at least one restriction enzyme is used to digest the PCR product and this restriction profile is different from that of the wild type sequence.