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For DNA extraction, physical and chemical processes are needed. The cellular contents are released by grinding the cell wall. A lysing solution is used to release the DNA from the sample. A detergent helps the DNA precipitate out of the solution, while sodium chloride causes the nucleic acid molecules to coalesce. Heat treatment is used to denature the DNA. Once the other cellular material is filtered off, and alcohol is added, the DNA can be collected using a spooling rod.

ABSTRACT

INTRODUCTION

DNA Sample Preparation1. To prepare a sample for DNA extraction, take 100 mL of distilled water and add to the enzyme bottle (Bromelain). Tighten the cap, shake well, and refrigerate.2. Put the ethanol in the freezer.3. To break open the cell wall, use mortar and pestle to grind the untoasted wheat germs into a fine powder.4. Mix 100 mL of lysing solution with the wheat germ powder into the beaker. For 10 min, stir and heat continuously at 60-65 degrees Celsius. Do not exceed past 65 degrees Celsius.5. Immediately cool to room temperature by placing the beaker in the ice bath after heating.6. Filter the mixture through 4 sheets of cheesecloth. Make sure the undissolved material does not transfer.

DNA Spooling1. Add 2 mL of the wheat germ filtrate to the test tube using a clean pipet.2. Add 2 mL of the enzyme solution into the test tube and gently mix the two for 5 min.3. Slowly lower the spooling rod into the lysate solution, letting it rest at the bottom of the tube.4. Add 4 mL of cold 70-95% alcohol to the lysate solution using a clean pipet to prevent cross contamination. Tip the tube at a 45-degree angle against the side of the tube to prevent the aqueous and the ethanol layers from mixing.5. Hold the tube at a 45-degree angle and rotate the spooling rod in a clockwise direction. Try not to touch the sides of the tube with the rod. Continue swirling the rod for several minutes until a visible mass of DNA has come out of the solution and attached to the rod.6. Air dry the extracted DNA for 5 min.

METHODS RESULTS CONCLUSIONS

After removing the spooling rod, air dry the extracted DNA for 5 min. When rotating the spooling rod, the DNA fibers come out of the solution and attach on to it. After the DNA has been extracted, it can be analyzed in many different ways, focusing on different genes, mutations, and proteins. When we removed our rod, we were able to see the DNA on the rod and some of the DNA inside of the tube. The DNA was yellow, wet, and a slimy mixture. Our pictures show the DNA on the rod and inside of the tube with the mixtures.

REFERENCES

Guha, Pokhraj et al. “A rapid and efficient DNA extraction protocol from fresh and frozen human blood samples.” Journal of clinical laboratory analysis vol. 32,1 (2018): e22181. doi:10.1002/jcla.22181. Accessed May 11, 2021.Lab Manual. Plant Biotechnology: DNA Extraction.Lõoke, Marko et al. “Extraction of genomic DNA from yeasts for PCR-based applications.” BioTechniques vol. 50,5 (2011): 325-8. doi:10.2144/000113672. Accessed May 11, 2021. Park, Jessica. “DNA Basics.” Understanding Genetics, 2017, genetics.thetech.org/ask-a-geneticist/two-genomes-cell. Accessed May 11 , 2021. Oppert, Brenda et al. “Optimized Extraction of Insect Genomic DNA for Long-Read Sequencing.” Methods and protocols vol. 2,4 89. 23 Nov. 2019, doi:10.3390/mps2040089. Accessed May 11, 2021.

ACKNOWLEDGEMENTS

Weeks Endowment.

DNA extraction is a process during which DNA is taken out of a cell using physical and chemical processes. When the cells are ground, the cellular contents are released. When the grounded material is placed into lysing solution, a detergent opens the nuclear membranes to release the DNA. Sodium chloride reacts with the negative phosphate ends of the DNA resulting in nucleic acid molecules to stick together. A citrate ion works as a chelating agent that helps hold together the magnesium ions. Both ions are needed by DNAase, an enzyme that breaks down DNA. Heat treatment at 65 degrees Celsius isused to deactivatethe enzyme. Anotherheat treatment at 80 degrees Celsius makes the DNA unfold. After the other cellular material, is filtered off, the filtrate is added to a test tube along with a protein-digesting enzyme that will remove extraneous proteins that were not denatured during the heat treatment. Coldalcohol is added to precipitate the DNA. After, a spooling rod is lowered into the solution. When the rod is rotated, the DNA fibers come out of the solution and attach to the rod. After the DNA has been extracted, it can be analyzed in many different ways, focusing on different genes, mutations, and proteins.

Baylor SchoolBrooklyn Gordon, Agda Laakso, Grace Phillips

DNA Extraction

Extracted DNA attached to the spooling rod after the extraction process.

This figure shows the amount of DNA that was extracted from the blood samples during the experiment. (Guha, Pokhraj et al).

This figure shows the amount of DNA that was extracted from the yeast samples and amplified using PCR. (Lõoke, Marko et al).

This figure shows the genomic DNA that was extracted from male T. castaneum.(Oppert, Brenda et al).