Dna Analysis : Strawberry Dna Extraction

As an eager fifth-grader in 2005, I quickly set my heart and sights on exploring genetics after a weeklong summer camp, “Designer Genes”, which ending in my running home with a small plastic tube of strawberry DNA I had proudly extracted, gushing to my family about everything I learned from transcription to translation. While new discoveries have drastically developed the field since, my interest and enthusiasm have yet to dwindle. As such, I elected to take advanced courses in biology/genetics and, beginning early as a rising senior in high school and ending only recently

that goes into creating a clone. First, scientist remove a somatic cell from an animal that they

Each human being has something called DNA. DNA is described as genetics and an extremely long macromolecule that is the main component of chromosomes and is the material that transfers genetic characteristics in all life forms. DNA constructs of two nucleotide strands coiled around each other in a ladder like arrangement with the sidepieces composed of alternating phosphate and deoxyribose units and the rungs composed of the purine and pyrimidine bases adenine, guanine, cytosine, and thymine. Each chromosome consist of one continuous thread-like molecule of DNA coiled tightly around proteins and contains a portion of the 6,400,000,000 basepairs that make up your DNA.

In order to extract DNA from any living thing, we needed to first gather the materials. Then we began the experiment. Step 1, put in a blender 1/2 cup of split peas (100ml), 1/8 teaspoon table salt (less than 1ml), and 1 cup cold water (200ml). Next, we blended the materials on high for 15 seconds. This allowed the pea cells to separate from each other, so we now had a really thin pea-cell soup. Step 2, poured our thin pea-cell soup through a strainer into another container. Added 2 tablespoons of liquid detergent (about 30ml) and swirled to mix. We then let the mixture incubate for 7 minutes. Poured the mixture into test tubes containers, and filled each about 1/3 full. Step 3, added a pinch of meat tenderizer to each test tube and stirred gently to make sure we didn’t break up the DNA. Step 4, tilted our test tube and slowly poured rubbing alcohol (70-95% isopropyl or ethyl alcohol) into the tube down the side so that it forms a layer on top of the pea mixture. Poured until we had about the same amount of alcohol in the tube as pea mixture. Alcohol is less dense than water, so it floats on top. From there we looked for clumps of white stringy stuff where the water and alcohol layers meet. The white stringy stuff was tangled DNA molecules. Thus, we completed DNA extraction. As we performed the experiment we made no changes to the original protocol.

Students will learn about the shape and function of DNA while extracting some from the cells of wheat germ.

Figure 1 Gel Electrophoresis for Replication Taster PTC. The gel is composed of an ethidium bromide stained 3% agarose gel demonstrating DNA fragments which were a depiction of PCR amplification. The agarose gel contains nine loading samples, including from left to right, the MW marker lane 1 precision mol mass standard, lane 2 TB undigested PTC (5µl of DNA, 5µl of master mix P, and 2.5µl of loading dye), lane 3 TB digested PTC (5µl of DNA, 5µl of master mix P, 2µl Fnu4HI, and 3µl of loading dye), lane 4 TB A(L)DH G (10µl DNA, 10µl master mix G, and 5µl loading dye), lane 5 TB A(L)DH A (10µl DNA, 10µl master mix A, and 5µl loading dye), lane 6 MG undigested PTC (5µl of DNA, 5µl of master mix P, and 2.5µl of loading dye), lane 7 MG digested PTC (5µl of DNA, 5µl of master mix P, 2µl Fnu4HI, and 3µl of loading dye), lane 8 MG A(L)DH G (10µl DNA, 10µl master mix G, and 5µl loading dye), lane 9 MG A(L)DH A (10µl DNA, 10µl master mix A, and 5µl loading dye).

The final experiment used E.coli so all we had to do to release the dna was to add the lysing solution since there is no cell wall to break. We used Iced ethyl alcohol to form a layer above the lysate solution for the dna to float into in all of the experiments as well as tipping the vial at a 45 degree angle while we spooled the dan on our patented glass dna extraction rod.

The gel was covered with a buffer and then six samples labeled A-F were deposited into the wells using a micropipette. Three of these samples (A, B, C) were control samples to compare to D, E, and F (the mother, child, and father). The safety cover was placed on the unit and then brought to a power source. The leads were connected the chamber and left for approximately 20 minutes. The agarose gel was removed from the tray and placed onto a sheet of plastic wrap. An Ethidium Bromide card was placed face down onto the gel to stain for approximately 5 minutes. Finally, the card was removed and the gel was placed on top of a UV light. The samples were pushed towards the center due to opposite electric charges. Agarose gel separates the DNA samples by the way they were cut. The restriction enzyme MST II cuts the DNA strand at CC/TNAGG where N is any nucleotide base. If the enzyme recognizes this, it is cut. If it does not recognize it then the strand is left whole. We were able to observe the DNA strands due to them being dyed and placed over a UV light. The control samples were utilized so that the other samples could be compared to test for their genotype. The data was analyzed in this way to differentiate between the different genotypes and the number of bars they

Plants are organisms that can reproduce sexually through meiosis and create more cells through mitosis (Russell et al. 2013). For studying mitosis, the common onion is an ideal choice. Because it is easy for onions to germinate without soil, it is easy to control any substances provided to the plant. The onion root tips are only a few cells thick and grow quickly making them ideal for time efficiency. The onion root tip needs to be squashed between the cover slip and the microscope in order to reduce the slide preparation’s total depth. To dye condensed chromosomes, such as those undergoing mitosis, a stain is used to make

(1) Carolina Biological Supply Company, 2004, Information, Strawberry DNA Extraction, 28th of January 2017, https://www.cpet.ufl.edu/wpcontent/uploads/2012/10/strawberry_dna_extract_kit.pdf

The results of the experiment were compared. As seen in the observations above, most data seemed to imply that there is more DNA in the pea sample. At the end of the experiment, three layers were visible, one was an alcohol layer at the top, a DNA precipitate and a suspension at the bottom (green homogenate for peas and yellow for onion). After comparing the key findings, it was determined that the hypothesis made before the experiment was incorrect. Cells with more chromosomes have more DNA but it is not noticeable visually. The amount of DNA that can be seen visually is dependent on the cell volume. Peas have a small cell volume as they have a small amount of water in the cytoplasm, which produces a lot DNA (“Genetic Science Learning Center”

Making the onion tip root cell slide was successful. Our results supported the hypothesis because we saw cells in the onion root tip in prophase, metaphase, and anaphase. As we went up in power objectives, each phase of the cell became more definitive. The cell root was a great indicator of the structures of the different cycles of the cell. This is important because we will be prepared for future labs working with the microscopes and can now adjust it for the best view of the slide. We practiced working with the compound light microscopes and different phases of the cell cycle. Onion root tips are useful to observe mitosis because the cells are frequently diving as the root grows. So when we stained the cell, we caught many cells in different phases. The significance of this lab was to better understand the process and stages of mitosis and meiosis and compare and contrast the mitotic process in plants and animals. We grasped the concepts of what the chromosomes look like, and what they look like in each step of the processes. Having read much about mitosis and meiosis, seeing these cells was the real application of describing and understanding the stages.

Much can be learned from studying an organisms DNA. The first step to doing this is extracting DNA from cells. In this experiment, you will isolate DNA from the cells of fruit. Materials (1) 10 mL Graduated Cylinder(2) 100 mL Beakers15 cm Cheesecloth1 Resealable Bag1 Rubber Band (Large. Contains latex pleasewear gloves when handling if you have a latex allergy).Standing Test TubeWooden Stir StickFresh, Soft Fruit (e.g., Grapes, Strawberries, Banana, etc.) ScissorsDNA Extraction SolutionIce Cold EthanolYou Must ProvideContains sodium chloride, detergent and waterFor ice cold ethanol, store in the freezer 60 minutes before use. Procedure If you have not done so, prepare the ethanol by placing it in a freezer for approximately 60 minutes.

The chemical and reagents used for the extraction and quantitation of DNA were: Plant DNAzol (0.3ml/0.1g), 100% ethanol (100%: 0.225 ml/0.1 g, 75%: 0.3 ml/0.1 g), Chloroform (0.3 ml/0.1 g), Plant DNAzol-ethanol solution: Plant DNAzol, 100% ethanol (1:0.75 v/v), TE buffer (10 mM Tris, 1 mM EDTA pH 8.0), 1.2% agarose gel (Agarose, 1X TAE buffer), 6X loading buffer (glycerol, Tris/EDTA pH 8.0, ethidium bromide), .25X TAE buffer, Restriction enzymes and Restriction endonuclease buffers. All the chemicals used were quality grade. The restriction

Due to the DNA’s specificity, samples can be utilised for identification. DNA is a nucleic acid composed of deoxyribose sugar bound to a phosphate group and one of four nitrogenous bases (adenine, guanine, cytosine and thymine). Each section of these three components are referred to as nucleotides, which are joined to the phosphate or sugar of another nucleotide by strong covalent bonds to form a backbone. The nitrogenous bases are joined to complimentary bases of another nucleotide (adenine with thymine, guanine with cytosine) to create a double stranded molecule (Figure 2). To complete the double helical structure, the molecule coils to compact it’s contents. DNA molecules can contain up to two million base pairs, with a human genome containing approximately 3 million base pairs. The random assortment of nitrogenous bases as well as the numerous mutations within certain DNA sequences, results in genetically diverese DNA molecules and genomes between individials.