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Extra info for Введение в теорию вероятностей и ее приложение. Том 2
Bhandal, N. , Usher-wood, P. N. , Barnard, E. , and Da&son, M. G. (1991) Sequence of a functional invertebrate GABA* receptor subunit which can form a chlmeric receptor wtth a vertebrate a subumt. EMBOJ. 10,3239-3245. t’ER 3 Analysis of Mitochondrial DNA Mutations Masashi Tanaka and Takayuki Ozawa 1. Introduction Human mitochondrial DNA (mtDNA) is a closed circular genome of 16,569 bp (I), encoding 13 subunits of four enzyme complexes (Complexes I, III, IV, and V) in the oxidative phosphorylation system (2).
18,438-439. 4. , and Gundelfinger, E. D. (1986) Primary structure ofa developmentally regulated mcotimc acetylchohne receptor protem from Droso@&z. EMBOJ 5, 1503-1508. 5. Barnard, E. , Darhson, M. , and Seeburg, P. (1987) Molecular biology of the GABA* receptor: the receptor/channel superfamily. Trends Neurosci. 10,502-509. 6. , Ruppersberg, J. , Schroter, K. , Geese, K. , and Pongs, 0. (1989) Molecular basis of functional diversity of voltagegated potassium channels m mammahan brain. EMBOJ 8,3235-3244.
1. First, the target sequence is amplified as a double-stranded DNA fragment by “symmetric PCR,” where primers Lr and H, are present in equal amounts. Primers L, and H, have the sequences identical to the light (L) and heavy (H) strands of mtDNA, respectively. Then, an excess of the H strand of the target region is generated by “asymmetric PCR,” where primer Hz is present in a large excess over primer FL,. Since the primer FL,, consists of both the M 13 universal sequence and the L strand sequence, the H strand synthesis starting from primer H, extends over primer FL,; therefore, the resulting single-stranded DNA incorporates the sequence complementary to the Ml3 universal sequence at its 3’ end.