BsmFI NEB#R0572

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	The precipitated amplification products were digested with the tagging enzyme BsmFI in a 50-μl reaction mixture containing 0.2 mM each dNTP, 1× NEB Buffer 4, 1× bovine serum albumin (BSA), and 2 U of BsmFI (NEB) for 1 h at 65°C. Restriction products were filled in by the addition of 3 U of T4 polymerase (NEB) to the reaction mixture and incubated for 20 min at 12°C. The reaction mixture was diluted to a volume of 200 μl with LoTE, and the products were extracted with PC8 and precipitated as described above.

Protocol tips

The precipitated amplification products were digested with the tagging enzyme BsmFI in a 50-μl reaction mixture containing 0.2 mM each dNTP, 1× NEB Buffer 4, 1× bovine serum albumin (BSA), and 2 U of BsmFI (NEB) for 1 h at 65°C. Restriction products were filled in by the addition of 3 U of T4 polymerase (NEB) to the reaction mixture and incubated for 20 min at 12°C. The reaction mixture was diluted to a volume of 200 μl with LoTE, and the products were extracted with PC8 and precipitated as described above.

Publication protocol

The precipitated amplification products were digested with the tagging enzyme BsmFI in a 50-μl reaction mixture containing 0.2 mM each dNTP, 1× NEB Buffer 4, 1× bovine serum albumin (BSA), and 2 U of BsmFI (NEB) for 1 h at 65°C. Restriction products were filled in by the addition of 3 U of T4 polymerase (NEB) to the reaction mixture and incubated for 20 min at 12°C. The reaction mixture was diluted to a volume of 200 μl with LoTE, and the products were extracted with PC8 and precipitated as described above.

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