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Methods for DNA Extraction Comparison

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  • Lisa Lyons

Abstract:

In this experiment, I extracted DNA from the cells of Green Split Peas and Chicken Livers. I used different variables for each. First, I did the experiment with all materials being cold. The second time I did the experiment with materials at room temperature. My objective was to see which method would extract more DNA. The results were that the materials being colder extracted more DNA than the room temperature materials. In either case, I was able to extract more from the Green Split Peas than the Chicken Livers both times.

Introduction:

DNA is short for deoxyribonucleic acid. Nucleic acid, which is the genetic material determining the makeup of all living cells and many viruses. It consists of two long strands of nucleotides linked together in a structure resembling a ladder twisted into a spiral. The rungs of the ladder are made up of nucleotides: adenine (A), thymine (T), cytosine (C), and guanine (G). DNA can replicate itself. DNA also serves as a template for synthesis of RNA in the presence of RNA polymerase. (APA, Dictionary. com)

DNA is located in the chromosomes in of the human body. It is like blueprints or instructions for hair color, eye color, height, pretty much everything. DNA can be used to identify criminals with unbelievable exactness when genetic evidence exists. Similarly, DNA can be used to clear suspects and clear persons wrongly accused or convicted of crimes. In all, DNA is ever-increasingly crucial to ensuring precision and fairness in the criminal justice system. (The United States Department of Justice) DNA is used in a few ways to solve crimes. Comparing a suspects DNA to DNA found at the scene, or it can be put in the database to try to find the offender's match. DNA can also be used to identify victims.

Materials and Methods:

  • Chicken Livers
  • Green Split Peas
  • 91% Rubbing Alcohol
  • Meat Tenderizer
  • Salt
  • Blender
  • 4 small glass Pyrex bowels
  • Small strainer
  • Wooden Chopstick
  • Measuring cup
  • Liquid soap

20150412_172421.jpg

Put in a blender:

  • 1/2 cup of split peas (with the chicken livers I used 3 medium sized livers. )
  • 1/8 teaspoon salt
  • 1 cup cold water

Blend on high for 15 seconds

Pour through a strainer into measuring cup.

Add 2 tablespoons liquid soap and swirl to mix.

Let the mixture sit for 5-10 minutes.

Pour the mixture into glass bowls, each about 1/3 full.

Add a pinch of meat tenderizer and swirl very carefully

Tilt bowls and pour in rubbing alcohol. (Until it forms a layer on top of mixture)

Results:

The first experiment I used very cold materials, with the peas and with the chicken livers. After blending the peas, I got a very light green colored water solution. After straining, there was a lot of shell like material at the bottom of the blender. I then added the 2 tablespoons of liquid soap to the mixture and allowed it to sit for about 10 minutes. I then transferred the "soup" into the Pyrex bowls, where I added a pinch of meat tenderizer and stirred very gently with the wooden chopstick. After stirring the meat tenderizer in, I then added the rubbing alcohol until it formed a layer on top of the mixture. Almost immediately, I was able to see the white colored stringy material in the peas. These experiments also produced 3 layers: debris at the bottom, water in the middle, DNA floating at the top. I was able to transfer the DNA to another Pyrex bowl with alcohol in it to get a better look.

unnamed.jpg20150412_165828.jpg

After blending the livers, I got a very reddish, beige colored water solution. After straining, there was liver material at the bottom of the blender. I then added the 2 tablespoons of liquid soap to the mixture and allowed it to sit for about 10 minutes. I then transferred the "soup" into the Pyrex bowls, where I added a pinch of meat tenderizer and stirred very gently with the wooden chopstick. After stirring the meat tenderizer in, I then added the rubbing alcohol until it formed a layer on top of the mixture. Almost immediately, I was able to see the white colored stringy material in the livers. These experiments also produced 3 layers: debris at the bottom, water in the middle, DNA floating at the top I was able to transfer the DNA to another Pyrex bowl with alcohol in it to get a better look.

The second experiment I used almost room temperature materials with the peas and with the chicken livers. After blending the peas, I got a very light green colored water solution. After straining, there was a lot of shell like material at the bottom of the blender. These experiments also produced 3 layers: debris at the bottom, water in the middle, DNA floating at the top. I then added the 2 tablespoons of liquid soap to the mixture and allowed it to sit for about 10 minutes. I then transferred the "soup" into the Pyrex bowls, where I added a pinch of meat tenderizer and stirred very gently with the wooden chopstick. After stirring the meat tenderizer in, I then added the rubbing alcohol until it formed a layer on top of the mixture. This time there was hardly any of the white stringy material I found in the first experiments, and barely anything to transfer.

20150412_171705.jpg

After blending the livers, I again got a very reddish, beige colored water solution. After straining, there was a lot of material at the bottom of the blender. I then added the 2 tablespoons of liquid soap to the mixture and allowed it to sit for about 10 minutes. I then transferred the "soup" into the Pyrex bowls, where I added a pinch of meat tenderizer and stirred very gently with the wooden chopstick. After stirring the meat tenderizer in, I then added the rubbing alcohol until it formed a layer on top of the mixture. These experiments also produced 3 layers: debris at the bottom, water in the middle, DNA floating at the top. This time there was hardly any of the white stringy material I found in the first experiments, and yet again, barely anything to transfer.

Discussion:

My results were surprising. These experiments also produced 3 layers: debris at the bottom, water in the middle, DNA floating at the top.

It really made a difference in the outcome just by using colder materials. Using ice-cold water and ice-cold alcohol will increase the yield of DNA. "Low temperatures protect the DNA by slowing down the activity of enzymes that could break it apart. The cold alcohol helps the DNA precipitate (solidify and appear) more quickly. Why would a cell contain enzymes that destroy DNA? These enzymes are present in the cell cytoplasm (not the nucleus) to destroy the DNA of viruses that may enter our cells and make us sick. A cell's DNA is usually protected from such enzymes (called DNases) by the nuclear membrane, but adding detergent destroys that membrane. " (gslc. genetics. utah. edu)

Out of the two, the warmer water experiments eliminated the most debris, but were not very successful at extracting as much DNA material.

References:

American Psychological Association: dna. (n. d. ).  Dictionary. com Unabridged. Retrieved April 12, 2015, from Dictionary. com website:http://dictionary. reference. com/browse/dna

Explorable : https://explorable. com/who-discovered-dna

Learn. Genetics, The Genetic Science Learning Center, Retrieved from: http://learn. genetics. utah. edu/content/labs/extraction/howto/faq/


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