DNA+exp+group1

After reading through the experiment booklet, create an easy to follow running sheet (dot point) of the key steps involved. You might also want to include some important notes on safety or errors to avoid. This must be done by Friday morning before class.

Link back to DNA manipulation SAC page

Rehydration of the DNA 1. Takes approximately 10-15 mins 2. Add distilled water to the DNA until it becomes a solution 3. USE GLOVES AS DO NOT WANT TO CONTAMINATE WITH OUR OWN DNA!

Preparation of DNA fragments 1. Add different restriction enzymes to different labelled micro tubes 2. Incubate to speed up the enzymes and cut the DNA faster 3. ONCE AGAIN USE GLOVES SO TO NOT CONTAMINATE DNA!

Preparation of aragose gel 1. Gel already prepared, add 10 mL of molten aragose to gel into the gel box so that it fills the cavity and is under the teeth of the comb 2. Leave the box and let gel set until solid 3. Cut 2 pieces of carbon paper and add to either end of gel box, these act as electrodes

Loading the gel 1. Pour approximately 10 mL of the buffer solution into the gel box, so it covers the just flows over the gel. Make sure the solution gets into the area where the electrodes are or they become useless. 2. Gently ease the comb out of the gel so to allow the buffer solution to slowly fill the wells 3. Leave the gel and solution to set 4. Add 2 microlitres of die to each DNA solution using a clean tips on the pipette to avoid contamination and mix well 5. Place the DNA solution in the well using a new tip so no contamination occurs. Don’t forget to remember which sample is place in which well

Running the gel 1. Attach wires to each electrode ensuring that the positive terminal is connected to the opposite side to where the wells are, this is because DNA runs from the negative to positive terminal. 2. Check that the electrodes are in contact with the buffer solution 3. Leave the gel to run for approximately 3.5 hours 4. Once 3.5 hours is up and the blue loading dye has reached the end disconnect the power source. Then rinse clips and dry