Eco32I (EcoRV) (10 U/µL)

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	The test for the presence of cohesive ends of the phage genome was performed as previously described (26), using the restriction enzymes HindIII, XhoI, Eco32I, SalI, and SmaI (Thermo Scientific). Briefly, phage DNA samples were divided into two parts after restriction digestion. The first part was loaded on the gel at 0°C, whereas the second part was heated at 70°C for 10 min prior to loading. Subsequently, all the samples were immediately separated by electrophoresis, and the DNA band patterns were compared.

Protocol tips

The test for the presence of cohesive ends of the phage genome was performed as previously described (26), using the restriction enzymes HindIII, XhoI, Eco32I, SalI, and SmaI (Thermo Scientific). Briefly, phage DNA samples were divided into two parts after restriction digestion. The first part was loaded on the gel at 0°C, whereas the second part was heated at 70°C for 10 min prior to loading. Subsequently, all the samples were immediately separated by electrophoresis, and the DNA band patterns were compared.

Publication protocol

"Typically, 0.3 μg of phage DNA was digested with 10 U of a restriction endonuclease (REase) for 2 h in a 20-μl reaction volume under conditions recommended by the manufacturer. The digestion products were analyzed by 0.7% agarose gel electrophoresis using a 10-kb Gene-ruler DNA ladder (Thermo Scientific) as a molecular size standard.

The test for the presence of cohesive ends of the phage genome was performed as previously described (26), using the restriction enzymes HindIII, XhoI, Eco32I, SalI, and SmaI (Thermo Scientific). Briefly, phage DNA samples were divided into two parts after restriction digestion. The first part was loaded on the gel at 0°C, whereas the second part was heated at 70°C for 10 min prior to loading. Subsequently, all the samples were immediately separated by electrophoresis, and the DNA band patterns were compared."

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Eco32I (EcoRV) (10 U/µL) below.

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