US 7,320,866 B2
Discrimination method of target base in DNA, and allele specific primer used in the method of the same
Hidenobu Yaku, Osaka (Japan); Hiroaki Oka, Osaka (Japan); and Tetsuo Yukimasa, Kanagawa (Japan)
Assigned to Matsushita Electric Industrial Co., Ltd., Osaka (Japan)
Filed on Dec. 04, 2006, as Appl. No. 11/607,900.
Claims priority of application No. 2006-156324 (JP), filed on Jun. 05, 2006.
Prior Publication US 2007/0281340 A1, Dec. 06, 2007
Int. Cl. C12Q 1/68 (2006.01); C12P 19/34 (2006.01)
U.S. Cl. 435—6  [435/91.1] 7 Claims
 
1. A method for discriminating a target base carried by a target DNA, comprising:
(1) a DNA elongation step comprising
(a) binding to said target DNA a test allele specific primer,
wherein said target DNA comprises the sequence 3′-(T-G-S1)-5′, S1 being the target base, and
wherein said test allele specific primer comprises the sequence 5′-(B3-B2-S1′)-3′, (B3, B2) being bases selected from the group consisting of (G, T), (G, G), (C, T), and (C, G),
wherein S1′ is the 3′ terminal base of the test allele specific primer and is complimentary to the base predicted to be S1; and
(b) causing a test DNA elongation reaction; and
(2) a discrimination step comprising measuring the efficiency of said DNA elongation to discriminate that said target is the same as said predicted base, or that said target base is distinct from said predicted base.