Patent ID: 7282126

Claim:
A method of determining the genotype of a sample polynucleotide having at least a first variant site, comprising: amplifying at least a portion of the sample polynucleotide to obtain first amplicons, the first amplicons including the first variant site; combining the first amplicons with first and second different polynucleotide controls, the first and second polynucleotide controls differing by at least one base therealong, the position of the at least one differing base corresponding to the first variant site of the sample polynucleotide; preparing a plurality of first duplexes, each of at least some of the first duplexes comprising (i) a polynucleotide strand of one of the first amplicons and (ii) a complementary polynucleotide strand of the first polynucleotide control; preparing a plurality of second duplexes, each of at least some of the second duplexes comprising (i) a polynucleotide strand of one of the first amplicons and (ii) a complementary polynucleotide strand of the second polynucleotide control; subjecting the first and second duplexes to temperature gradient electrophoresis (TGE) to obtain first and second electrophoresis data; and determining the genotype of the first variant site of the sample polynucleotide based on the first and second electrophoresis data.