Patent ID: 6509160
Filing Date: 2003-01-21
Classification: C12Q

Abstract:
A method for identifying differences between a first and second nucleic acid sample, comprising:fragmenting the samples with a first type IIs endonuclease;(a) ligating to the fragments a first adapter sequence comprising a second type-IIs endonuclease recognition site; (b) fragmenting the samples with a third endonuclease and ligating a second adapter sequence to the fragments; (c) amplifying the fragments; (d) marking the amplified fragments from the first sample; (e) mixing the first and second samples under conditions that will allow heteroduplex formation between sequences in the first and second samples; (f) selectively degrading one strand from heteroduplexes which contain a mismatch and isolating the remaining single-stranded fragments; (g) creating double-stranded DNA from the single-stranded fragments and amplifying the double-stranded DNA; (h) cleaving the DNA with the second type-IIs endonuclease; and determining the sequence of polynucleotides between the first and second type-IIs endonuclease recognition sites.