VpaK11BI (AvaII) restriction enzyme

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	After completing duplex PCR, PRA using 2 restriction enzymes, AvaII (TaKaRa) and HaeIII (TaKaRa), was performed to identify the 105 NTM strains that produced 515-bp hsp65 DNA. PRA reactions were performed as follows. Ten microliters of PCR products was transferred to a fresh microcentrifuge tube and digested with restriction enzymes according to the manufacturer's instructions. Following digestion, mixtures were electrophoresed on 3% agarose gel gels.

Protocol tips

After completing duplex PCR, PRA using 2 restriction enzymes, AvaII (TaKaRa) and HaeIII (TaKaRa), was performed to identify the 105 NTM strains that produced 515-bp hsp65 DNA. PRA reactions were performed as follows. Ten microliters of PCR products was transferred to a fresh microcentrifuge tube and digested with restriction enzymes according to the manufacturer's instructions. Following digestion, mixtures were electrophoresed on 3% agarose gel gels.

Publication protocol

A PRA algorithm for differentiating mycobacteria was constructed using MapDraw (version 3.14; DNASTAR, Madison, Wis.). After completing duplex PCR, PRA using 2 restriction enzymes, AvaII (TaKaRa) and HaeIII (TaKaRa), was performed to identify the 105 NTM strains that produced 515-bp hsp65 DNA. PRA reactions were performed as follows. Ten microliters of PCR products was transferred to a fresh microcentrifuge tube and digested with restriction enzymes according to the manufacturer’s instructions. Following digestion, mixtures were electrophoresed on 3% agarose gel gels.

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Manufacturer protocol

Download the product protocol from Takara Bio Inc for VpaK11BI (AvaII) restriction enzyme below.

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