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. |
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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.
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