Enzyme digest and treatment pJH1 with SmaI, and Antarctic Phosphatase

3/13/2015

1-In a 1.5mL tube combine the following:

• DNA     15 μL (1-5 mg) 

• Restriction Enzyme(1μL SmaI )

• Buffer_cutsmart_( 2 μL )

• dH2O ( 2 μL )

              20 μL total.Incubate 25 °C    for 2 hours, inactivation 65  °C   for 20 minutes.
2-Antarctic Phosphatase catalyzes the removal of 5´ phosphate from DNA.

• DNA     15 μL (from #1 above Δ SmaI) 

• Antarctic Phosphatase(1μL SmaI )

• Buffer ( 2 μL )

• dH2O ( 2 μL )

              20 μL total.Incubate 37 °C    for 15 minutes, inactivation 70  °C   for 5 minutes.