BfaI NEB#R0568

Protocol tips

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	Restriction digestion of 1 μg of custom-designed DNA was carried out at 37°C for 4 h using 30U AluI, 80U BfaI, 30U HaeIII, 15U HpyCH4V, 10U MluCI, 10U MseI, and 10U MspI. Restriction products were size separated in 2% agarose.

Protocol tips
Restriction digestion of 1 μg of custom-designed DNA was carried out at 37°C for 4 h using 30U AluI, 80U BfaI, 30U HaeIII, 15U HpyCH4V, 10U MluCI, 10U MseI, and 10U MspI. Restriction products were size separated in 2% agarose.

Publication protocol

Synthetic DNA bearing the seven restriction sites approximately every 50 nt was obtained from Integrated DNA Technologies, Inc. (Coralville, IA, USA). Restriction enzymes were obtained from New England Biolabs (Ipswich, MA, USA). The synthetic insert was amplified by PCR using the following cycle parameters: initial 95°C incubation, 30 cycles of 95°C/10 s denaturing, 66°C/10 s annealing, and 72°C/20 s extension, and final 72°C for 7 min incubation. Primer pairs used are shown in Figure 1. Unique band amplification was confirmed by gel electrophoresis and PCR products were purified using the MinElute PCR kit (Qiagen). Restriction digestion of 1 μg of custom-designed DNA was carried out at 37°C for 4 h using 30U AluI, 80U BfaI, 30U HaeIII, 15U HpyCH4V, 10U MluCI, 10U MseI, and 10U MspI. Restriction products were size separated in 2% agarose.

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