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- W1565360419 abstract "Aims: The aim of the present study was to rapidly optimize enterobacterial repetitive intergenic consensus (ERIC)-PCR amplification systems for fingerprinting rat’s intestinal microflora. Methods and Results: Orthogonal array design and statistic analysis methods were attempted to rapidly optimize ERIC-PCR reaction system for fingerprinting intestinal microflora. The results showed that variations of the four factors (Mg2+, dNTP, primer and HotstarTaq polymerase concentrations) changed the fingerprinting patterns significantly. The order of effects of those factors on fingerprinting patterns was primers (F = 274·000, P = 0·000), Hotstar Taq polymerase (F = 197·000, P = 0·001), Mg2+ (F = 181·000, P = 0·001) and dNTP (F = 27·000, P = 0·011). The optimal ERIC-PCR condition was containing 200 μmol l−1 dNTP, 2·5 mmol l−1 Mg2+, 0·4 μmol l−1 primer, 1 U HotstarTaq DNA polymerase namely 25 μl reaction system, which is proved to be a simple, fast and reliable method suitable for fingerprinting rat’s intestinal microflora. Conclusions: The results suggest that Mg2+, dNTP, primer and HotstarTaq polymerase concentrations play important roles on ERIC-PCR fingerprinting patterns. Orthogonal array design is a considerable method to optimize ERIC-PCR reaction system for its rapidness, simplicity, potential to investigate mutual effects of parameters. Significance and Impact of the Study: It is the first report on optimization of ERIC-PCR amplification systems for fingerprinting intestinal microflora using orthogonal array design or statistic analysis methods and systematically observing the effects of variables of reaction conditions." @default.
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- W1565360419 date "2007-07-30" @default.
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- W1565360419 title "Orthogonal array design in optimizing ERIC-PCR system for fingerprinting rat’s intestinal microflora" @default.
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- W1565360419 doi "https://doi.org/10.1111/j.1365-2672.2007.03440.x" @default.
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