Restriction Enzymes

What are they? Are enzymes that cleave the DNA in specific locations. Also called endonucleases “Restriction Sites” = the specific place on the DNA molecule Purpose of today’s Lab: 1. perform a digest of DNA using three different restrictions enzymes= EcoRI, BamHI, HindIII 2. demonstrate proper technique using a micropipette 2. describe the type of ends a restriction enzyme leaves

Micropipetting Very expensive instrument for measuring Measures very small volumes: microliters (ul) Three types: P-1000 P-200 P-20

Parts of the Micropipette Tip Plunger Adjustment dial Volume window

Using the Micropipette Detemine the volume to be pipette. Rotate the dial to the appropiate setting. NEVER GO ABOVE THE MAXIMUM VOLUME THAT IS MADE FOR THE MICROPIPETTE Use a clean pipette tip from box. Gently push micropipette onto tip. To draw up solution – depress plunger to first stop; release plunger and draw up solution; place tip at bottom of tube; depress plunger to second stop. Eject the used tip over and into the waste basket on table.