FastDigest Eco91I

Protocol tips

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	Amplified fragments were digested with HaeIII and Eco91I (BstEII) restriction enzymes (FastDigest), under conditions recommended by the manufacturer (ThermoScientific), separated by electrophoresis in 4% agarose gels, and visualized by staining with ethidium bromide (0.5 μg/mL) and exposure to UV light (λ = 320 nm).

Protocol tips
Amplified fragments were digested with HaeIII and Eco91I (BstEII) restriction enzymes (FastDigest), under conditions recommended by the manufacturer (ThermoScientific), separated by electrophoresis in 4% agarose gels, and visualized by staining with ethidium bromide (0.5 μg/mL) and exposure to UV light (λ = 320 nm).

Publication protocol

"For the amplification of a 441 bp fragment of the hsp65 gene Tb11 and Tb12 primers were used, as described by Telenti et al. [15]. The PCR mixtures were prepared with a TopTaq Master Mix kit (Qiagen) in a final volume of 50 μL containing ca. 10 ng of genomic DNA. After initial denaturation at 94°C for 3 min, the reaction mixture was run through 35 cycles of denaturation at 94°C for 30 s, annealing at 57°C for 30 s, and extension at 72°C for 30 s, followed by a final extension at 72°C for 10 min. Amplified fragments were digested with HaeIII and Eco91I (BstEII) restriction enzymes (FastDigest), under conditions recommended by the manufacturer (ThermoScientific), separated by electrophoresis in 4% agarose gels, and visualized by staining with ethidium bromide (0.5 μg/mL) and exposure to UV light (λ = 320 nm).

Strains were classified into subtypes based on their PCR-REA patterns obtained in two separate PCR-REA assays."

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for FastDigest Eco91I below.

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