SNP Flashcards
Identifying a SNP with an RFLP assay
Firstly the DNA containing the target gene/SNP is amplified using an upstream and downstream primer. A restriction endonuclease is added to digest the fragment depending on its palindromic sequence at 37 degrees C. The fragments are run on gel electrophoresis to display fragment size and number.
A wild type individual that has no mutation will have one large fragment on the gel since the SNP has not cut since its sequence isn’t palindromic.
A homozygous recessive individual for the mutation will have two smaller fragments on the gel, as the sequence at the SNP is palindromic so is cut.
An individual that is heterozygous for the mutation (one dominant and one recessive allele) exhibits three bands on the gel - the large band from the dominant allele and two smaller bands from the recessive allele.
This method is cheap and efficient but has a low throughput.
