US 7,601,495 B2
Methods, compositions, and kits comprising linker probes for quantifying polynucleotides
Caifu Chen, Palo Alto, Calif. (US); Dana Ridzon, San Francisco, Calif. (US); Zhaohui Zhou, Fremont, Calif. (US); Kai Q. Lao, Pleasanton, Calif. (US); and Neil A. Straus, Emeryville, Calif. (US)
Assigned to Applied Biosystems, LLC, Carlsbad, Calif. (US)
Filed on May 31, 2005, as Appl. No. 11/142,720.
Application 11/142720 is a division of application No. 10/947460, filed on Sep. 21, 2004.
Prior Publication US 2006/0078906 A1, Apr. 13, 2006
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/68 (2006.01); C12P 19/34 (2006.01)
U.S. Cl. 435—6  [435/91.2] 7 Claims
 
1. A method for quantifying a first small nucleic acid in a reaction mixture comprising the first small nucleic acid and a second small nucleic acid, wherein the first small nucleic acid and the second small nucleic acid differ by only two nucleotides, wherein the small nucleic acids are 18-25 nucleotides in length, said method comprising;
selectively forming an extension reaction product of the first small nucleic acid, wherein a 3′ target-specific portion of a linker probe hybridizes to the first small nucleic acid but not to the second small nucleic acid, wherein the extension reaction product is created by extending from the 3′ target-specific portion of the linker probe;
exponentially amplifying the extension reaction product of the first small nucleic acid in a polymerase chain reaction (PCR); and
selectively quantifying the amplified extension reaction product of the first small nucleic acid over the second small nucleic acid by hybridizing a detector probe to a sequence corresponding to the first small nucleic acid.