Enzymes used

Reaction system

Reaction temperature

Incubation time

Msp I

PCR reaction mixture―10 μL

Nuclease-free water―10 μL

1× CutSmart™ Buffer―2.5 μL

Msp I (5U)―0.5 μL

37˚C

1 hour

Bsa HI

PCR reaction mixture―10 μL

Nuclease-free water―10 μL

1× NE Buffer 4―2.5 μL

Bsa HI (5U)―0.5 μL

60˚C

1 hour

Hinf I

PCR reaction mixture―10 μL

Nuclease-free water―10 μL

1× NE Buffer 4―2.5 μL

Hinf I (5U)―0.5 μL

37˚C

1 hour

Bst EII

PCR reaction mixture―10 μL

Nuclease-free water―10 μL

1× NE Buffer 3―2.5 μL

Bst EII (5U)―0.5 μL

60˚C

1 hour