Restriction Digest of DNA
Select restriction enzymes to digest your plasmid.
Determine an appropriate reaction buffer by reading the instructions for your enzyme.
In a 1.5mL tube combine the following:  DNA (all amounts are for a typical reaction; your amount may vary depending on the enzymes).
Typical mixture for single digest: Nuclease-free Water, 16 ul; 10X Buffer EcoRI2 ulDNA (0.5/1 ug/ul), 1 ul; EcoRI0. 5-2 ul;
Typical Mixture for Double Digest: Nuclease-free Water, 15 ul; 10X Buffer, 02 ul; DNA (0.5-1 ug/ul), 1 ul; EcoRI, 1 ul; PstI, 1 ul;
Mix gently by pipetting.
Incubate tube at appropriate temperature (usually 37°C) for 1 hour.
