1. Recombinant DNA Bacterial Transformation Student Instructions Plasmid Digest

2. Recombinant Transformation Plasmid Digest Obtain four 1.5 ml reagent tubes labeled: - pAmp - pKan - 2XRB”( Restriction Buffer ) - B/H” (BamHI and HindIII enzymes).

3. Use the matrix below as a checklist while adding reagents to the pAmp and pKan tube. Use a fresh pipette tip for each reagent. Add all reagents directly to the solution. Recombinant Transformation Plasmid Digest NOTE: The buffer is always added before the enzymes. Tube2X RBB/H __________________________________________________________ pAmp 7.5 µl2 µl pKan 7.5 µl2 µl

4. Recombinant Transformation Plasmid Digest Place the reaction tubes in a 37 o C water bath and incubate for 30-60 minutes.

5. Recombinant Transformation Plasmid Digest After incubation, freeze the reaction at -20 o C until you are ready to proceed to Ligation. You can store reactions at this temperature for several days.