BioBrick Restriction Digest

Reagents:

- dsH2O;

- 10x Buffer (1 – 2 – 3 – 4);

- 10x BSA;

- Restriction Enzymes (EcoRI, Xbal, SpeI, PstI);

- DNA.

Steps:

1. Make a master mix with everything except the DNA. Make sure that:

• The final concentration of BSA and buffer is 1x.

• 5 to 10 enzyme’s unit are necessary for 1 µg plasmidial DNA digest.

• Add to digest system the quantity of DNA enough for 0.5 – 1 µg. If the system will be purified from the agarose gel lately, add quantity of DNA for the digest system of 1 – 2 µg.

2. Pipet aliquots to each sample tube and add DNA.

3. Incubate at 37°C for 2 – 3 hours. Check the eletrophorectic profile.