SspI restriction enzyme

Protocol tips

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	5 μl of the PCR product and 2 μl of the restriction enzyme were prepared by mixing 2 μl of 10x buffer, and sterile free water was added for a total volume of 20 μl. The tubes were incubated at 37°C for approximately 4 h. The digested PCR products were then separated on 2.5% agarose gel.

Protocol tips
5 μl of the PCR product and 2 μl of the restriction enzyme were prepared by mixing 2 μl of 10x buffer, and sterile free water was added for a total volume of 20 μl. The tubes were incubated at 37°C for approximately 4 h. The digested PCR products were then separated on 2.5% agarose gel.

Publication protocol

To further discriminate between targeted animals, expected nucleotide sequences were restriction mapped with the Mapdraw program of DNASTAR (NY, USA). After testing, one restriction enzyme SspI (TakaRa) was selected to digest the PCR products in separate reactions. Briefly, 5 μl of the PCR product and 2 μl of the restriction enzyme were prepared by mixing 2 μl of 10x buffer, and sterile free water was added for a total volume of 20 μl. The tubes were incubated at 37°C for approximately 4 h. The digested PCR products were then separated on 2.5% agarose gel.

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