Patent ID: 6825010
Filing Date: 2004-11-30
Classification: C12Q

Abstract:
A method for identifying at least one target nucleotide within a target sequence comprising:generating multiple fragments of the target sequence by digesting the target sequence with at least one restriction enzyme; amplifying the multiple fragments using a first set of primers to generate multiple amplified fragments, wherein the first set of primers comprises a first primer and a second primer; amplifying at least one subset of the multiple amplified fragments to generate at least one amplified subset of the multiple amplified fragments using a first set of extended primers, wherein the first set of extended primers comprises a first extended primer which comprises the sequence of the first primer and one additional nucleotide on the 3â€² end and a second extended primer which comprises the sequence of the second primer and one additional nucleotide on the 3â€² end; amplifying the at least one subset of the multiple amplified fragments to generate at least one second subset of amplified fragments using a second set of extended primers, wherein the second set of extended primers comprises a third extended primer which comprises the sequence of the first extended primer and one additional nucleotide on the 3â€² end and a fourth extended primer which comprises the sequence of the second extended primer and one additional nucleotide on the 3â€² end; and identifying the at least one target nucleotide within a target sequence by identifying the at least one target nucleotide in at least one of the amplified subsets of the multiple amplified fragments.