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. , Esen , N. , Özkütük , A.A. : Comparison of polymerase chain reaction-restriction enzyme analysis method and DNA sequence analysis results in the identification of non-tuberculous mycobacteria

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resistance determinants were found in this study. Among 23 aac(6)-Ib , PCR positive strains BstF5I restriction enzyme analysis detected nine aac(6)-Ib-cr variant, and these were carried by E. coli, M. morganii, Klebsiella spp., and Enterobacter spp

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37 °C with 50 U of the XbaI restriction enzyme according to the manufacturer's instructions. The restriction fragments were separated through PFGE in 1% agarose gel (Bio-Rad, USA) with 0.5x TBE buffer (45 mM Tris, 45 mM boric acid, and 1.0 mM EDTA [pH

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, these isolates were cultured on trypticase soy agar and then prepared bacterial plugs lysed with lysis buffer. The plugs were digested with restriction enzyme XbaI (Fermentas, Lithuania) for about 150 min at 37 °C. Then, electrophoresis of the plugs was

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) in E. coli DH5α. The clones were verified by plasmid isolation and restriction enzyme digestion. The recombinant pET-28a(+) vectors carrying erp, hspR, lppX, mmaA4 , and ompA genes were introduced into E. coli BL21(DE3) competent cells (Novagen

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use of the Prime Script RT reagent Kit with gDNA Eraser (Takara, Japan) with following the operating instructions. The specific primers with cleavage sites (PstI and EcoRI) of restriction enzyme introduced, RcFATA-F (5′-AAAACTGCAGATGTTAAAAGTACCTTGTTG-3

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Biologia Futura
Authors: Muhammad Saad, Helen Mary, Umar Amjid, Ghulam Shabir, Kashif Aslam, Shahid Masood Shah, and Abdul Rehman Khan

restriction enzyme and the other subsample was digested with Hpa II by incubation at 37 °C overnight and enzymes inactivation at 80 °C for 20 min. The restricted samples were then ligated with Eco RI and Hpa II linkers (Table  1 ) using T4 DNA ligase enzyme

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