Science Fair Project Keep Calm and Split DNA Charisma Ware - Carleen McNees - Syracuse Junior High
Problem How can you separate DNA when they are to small to see, let alone cut?
Hypothesis If I put a group of DNA / RNA in a gel electrophoresis chamber then the DNA / RNA will separate because scientists use gel electrophoresis chambers to separate DNA and RNA all of the time.
Gel Electrophoresis Research In the early days of DNA manipulation, DNA fragments were laboriously separated by gravity. In the 1970s, the powerful tool of DNA gel electrophoresis was developed. This process uses electricity to separate DNA fragments by size as they migrate through a gel matrix Check out Gel Electrophoresis Chamber Video on the website below http://www.dnalc.org/resources/animations/gelelectrophores is.html Play the Gel Electrophoresis Game on the website below it teaches you how to use a Gel Electrophoresis Chamber and why we use them. http://learn.genetics.utah.edu/content/labs/gel/
Materials Plastic Ice Cube Tray Aluminum foil 9 volt battery Connecting wires with alligator clips Scissors Buffer solution (baking soda and water, mixed evenly) Medicine dropper Agar solution Scale Measuring Cup with ml Pot & stove DNA (Food coloring)
Variables Independent: DNA Food Coloring Dependent: Gel Electrophoresis Chamber Constant: Buffer Solution Control: N/A
Procedure- Part 1 Setting the Gel 1. Clean and dry two mold chambers of an ice cube tray. 2. Get the agar solution. 3. Pour this solution into each of the tray slots. 4. Leave alone or place the tray in a refrigerator to speed up the solidification process of the gelatin. Agar Solution: Mix 3g of Agar Powder with 260mL of Water. Bring to a boil and stir evenly. Let cool and follow steps 1-4.
Procedure Part 2 Building the Chamber 1.Make a blade-like tool to cut into the solid gel. 2. Use this blade to poke a slot-shaped well into the gelatin. The well should be in about the middle. Use this tool to poke, cut, and remove the gel material so that a thin (less than 1mm) rectangular slot is created. 3. Cut two strips of aluminum foil (about 1 cm by 4 cm) for the chamber electrodes. Insert these along opposite inner sides of the mold.
Procedure Part 3 Running the Sample 1. Fill to the surface with buffer solution. A layer of about 3 mm of buffer must cover the top of the gel and fill the rectangular well. 2. Fill your pipette or medicine dropper with green food coloring. Put the tip in the buffer and position it at the bottom of the well. Slowly release some of your sample and observe how if flows upward and fill the well. Continue releasing the sample until the well is filled. Carefully remove the tip of the pipette or dropper from the chamber, trying not to mess with the buffer solution. 3. Use alligator clips to connect a 9-volt battery to the tops of the aluminum foil tips. Wait one hour. 4. Check on the gel. NOTE: You can increase the speed of the separation by wiring up several 9-volt batteries together. 5. Reattach the clips and continue examining the gel at 30-minute intervals.
Observations All of the samples are mostly negatively charged. All of the secondary colors split into the primary colors that create them. Yellow traveled farther than red and blue in the green and orange samples.
Red (observations) After 1 hour of sitting undisturbed, the red sample had bubbles near the positive electrode which means that the red coloring is mostly negatively charged. The bubble were all red.
Orange (observations) After 1 hour of sitting undisturbed, the orange sample had bubbles near the positive electrode which means that the orange coloring is mostly negatively charged. The bubbles were red and yellow, the two colors I used to create the orange coloring. The yellow traveled farther than the red.
Yellow (observations) After 1 hour of sitting undisturbed, the yellow sample had bubbles near the positive electrode which means that the yellow coloring is mostly negatively charged. The bubble were all yellow.
Green (observations) After 1 hour of sitting undisturbed, the green sample had bubbles near the positive electrode which means that the green coloring is mostly negatively charged. The bubble were yellow and red. The yellow traveled the furthest.
Blue (observations) After 1 hour of sitting undisturbed, the blue sample had bubbles near the positive electrode which means that the blue coloring is mostly negatively charged. The bubble were all blue.
Purple (observations) After 1 hour of sitting undisturbed, the purple sample had bubbles near the positive electrode which means that the purple coloring is mostly negatively charged. The bubble were blue and red. The red traveled farther than the blue.
Conclusion You can separate DNA with a Gel Electrophoresis Chamber. My hypothesis was correct. I would like to try this experiment again but, next time I would get more than two 9 volt batteries and make more colors.