DAY TWO- DNA digestion and gel preparation.
What do restriction enzymes do?
How does the DNA of a child compare to its parents?
Put the steps in the proper order to digest the DNA samples using the restriction enzyme, HindiIII. You will use 5 uL of DNA for each sample. Get your procedure approved by the teacher before you begin. Remember to use fresh pipette tip for each transfer!
_____ Add 2 uL buffer to each tube. Add 5 uL of the corresponding DNA to each tube.
_____ Put on goggles, gloves, and assemble all equipment
_____ Label 6 microfuge tubes M1+, M1-, B1+, B1-, S+, S- (+ will receive HindIII enzyme, - will not)
_____ Add 1 uL HindIII to all tubes receiving the enzyme (those labeled +)
_____ Add sterile H2O to bring total volume in tube to 20 uL
_____ Incubate at 37°C for 30 minutes in the incubator block.
_____ Place all samples in the microcentrifuge and spin for five seconds to ensure all liquid is at bottom of tube.
Preparation of Gel-
How would you prepare 50 mL of a 1% agarose solution? (the solvent will be the 1XTAE buffer prepared previously.) Show a correct mathematical setup, and write out the steps.