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Once we verified that marginal posterior distributions of the two runs had achieved similar solutions, a total of 150,000 genealogies were analysed by using the nested models option in the "Load Trees Mode" (L-Mode).
Borrelia species were first identified by using the nested PCR reported by Ras et al. (4 ).
However, by using the nested ANOVA-design it was possible to discriminate between different sources of variation.
To determine molecular markers of resistance, we amplified and sequenced the PF3D7_1343700 gene by using the nested PCR method, as described (3 ), with some modifications.
Each PCR product was eluted from the filter paper into 50 μL of RNase-free water and reamplified by using the nested primer set.
S-amplified product (portion of nonstructural protein) was obtained by using the nested RT-PCR technique published by Sall et al. (12 ).
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Nucleic acids were extracted from 1-mL plasma volumes (MagNA Pure LC, Roche Diagnostics, Meylan, France) (2 ) and tested for KIs-V DNA by using the same nested PCR system for screening Japanese blood donors (1 ).
The full length sequences of 1Ay ORFs were obtained by using the method of nested deletion.
The first-round PCR products were produced by using the first stage of the nested PCR protocol; in the second round, the P. malariae specific primers of Padley et al. (7 ), which lie wholly within the first-round PCR product, were used to amplify a final product of 425 bp.
Here, we consider the continuous-discrete EKF and show that its quality is raised by using the adaptive sixth-order nested implicit Runge Kutta (NIRK) method of Gauss type with automatic local and global error controls.
The purpose of our study was to assess the occurrence of these 2 species in different types of endodontic infections by using the sensitive 16S rDNA based nested polymerase chain reaction approach.
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