Patent ID: 6576448
Filing Date: 2003-06-10
Classification: C12Q,Y10T

Abstract:
A method of circularizing and amplifying large fragments of DNA comprising:(a) providing a DNA-adapter construct which has a DNA fragment ligated to an adapter, wherein the adapter comprises first, second, and third oligonucleotides, wherein the first and the second oligonucleotides are annealed to the third oligonucleotide, wherein a gap is present between the first and the second oligonucleotides annealed to the third oligonucleotide, and wherein the second oligonucleotide has a 5â€² region present at the gap that is noncomplementary to the third oligonucleotide, and wherein first and second ends of the DNA fragment are ligated to first and second ends of the adapter to form a circle; and (b) amplifying the DNA-adapter construct using DNA pol III in a rolling circle amplification.