US 7,320,860 B2
Nucleic acid amplification method
Ulf Landegren, Uppsala (Sweden); Mats Gullberg, Uppsala (Sweden); and Mats Nilsson, Sigtuna (Sweden)
Assigned to Olink A.B., Uppsala (Sweden)
Appl. No. 10/483,900
PCT Filed Jul. 12, 2002, PCT No. PCT/SE02/01378
§ 371(c)(1), (2), (4) Date Jul. 20, 2004,
PCT Pub. No. WO03/012119, PCT Pub. Date Feb. 13, 2003.
Claims priority of application No. 0118959.6 (GB), filed on Aug. 03, 2001.
Prior Publication US 2005/0287526 A1, Dec. 29, 2005
Int. Cl. C12Q 1/68 (2006.01); C12P 19/34 (2006.01)
U.S. Cl. 435—6  [435/91.1; 435/91.2] 15 Claims
 
1. A method of analyzing a circularized nucleic acid, comprising amplifying the circularized nucleic acid to provide an amplification product which comprises a concatemer of a sequence to be analyzed, the method further comprising:
(i) monomerizing said amplification product using a restriction enzyme and an oligonucleotide complementary to the amplification product which is present in excess over the number of monomers contained in the amplification product, wherein the restriction enzyme cleaves any amplification product-oligonucleotide hybrids;
(ii) circularizing said monomers derived from the amplification product by hybridization to uncleaved oligonucleotide remaining from step (i) and ligation;
(iii) amplifying said circularized monomers in a rolling-circle amplification reaction;
(iv) optionally repeating steps (i) to (iii); and
(v) detecting or analyzing the amplification products.