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KMID : 0364820040400030254
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Korean Journal of Microbiology
2004 Volume.40 No. 3 p.254 ~ p.256
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Treatment of Genomic DNA with Restriction Enzyme(s) Improves Amplification Efficiency by Polymerase Chain Reaction
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민해기/Min HK
장영효/Chang YH
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Abstract
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Polymerase chain reaction (PCR) is a powerful tool for precisely amplifying selected DNA sequences that have had a broad impact on genomic studies. When examining human [alpha]- and [beta]- tryptase genes which have 95% DNA homology, inconsistent PCR amplification of genomic sequences hampered our progress. This study suggests that long PCR technique on the original DNA digested with restriction enzymes improves both efficiency and sensitivity of PCR. These improved results seem to derived from the effective denaturation of the original genomic DNA template or reduction of formation of secondary structures that block either primer annealing or extension in PCR. Elimination of homo- or hetero-duplex products derived from highly homologous genes provides an additional advantage in this study. This communication describes how the use of restriction enzymes improved these efficiencies, and also facilitated studies of highly homologous genes including tryptase genes.
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KEYWORD
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