June 4, 2012

Deoxyribonucleic Acid Extraction from Commercial Strawberries

Priyank Patel

Course Code: SBI4UPeriod 2 Student Number: 310059860 Submitted to: Mr. Jones

Purpose: To extract a significant amount of DNA from commercially available strawberries (Fragaria cv.) and to examine the yield under a microscope. Materials: Mortar and pestle Coffee filter paper Funnel 250 mL beaker Stirring Rod Test tub and test tube holder Procedure: 1. 2. 3. 4. 5. 6. 7. The strawberry was placed in the mortar. The strawberry was grinded using the pestle, into a consistent pulp. 10mL of the extraction fluid was added to the pulp, and was grinded further. Filter paper was folded into a cone and placed in a funnel over the beaker. The strawberry pulp was poured into the folded cone and left to filter for 10 minutes. 15mL of the cold ethanol was placed into a test tube. Using a pipette, some of the strawberry filtrate was placed into the test tube with the cold ethanol. 8. The test tube was swirled a couple of times, and placed back on the test tube holder/rack. 9. The white precipitate formed was observed. 10. Using the stirring stick, a small sample of the white precipitate was removed and used to prepare a wet mount slide. 11. The wet mount slide was placed under a microscope and observed. Apparatus Diagram: Retort stand and ring clamp 10 mL solution of dish soap and NaCl 15 mL of ethanol at 4 degrees Celsius 2 large strawberries Wooden Stick

Observation:

Figure 1: Test tube with white precipitate

Figure 2: View of wet mount slide through microscope.

Discussion of Results and Observations: The above pictures are of the lab as it was being performed. Figure 1 was taken after the strawberry filtrate was added to the test tube with cold ethanol. At the top and near the middle, the white swirly precipitate is the isolated DNA. It could be seen that the DNA was white and stringy. The bottom of the test tube contained other cellular components along with strawberry pigment. Figure 2 is of a wet mount slide of the DNA extracted under a microscope. When the extraction solution was added, the detergent dissolved the cell and nuclear membranes. This allowed it to be easier to access the highly protected DNA. Because the membranes are made of lipids (fat), the detergent will cut through the membrane. Then the salt (NaCl) isolated the DNA and proteins. Finally the cold ethanol clumped together the DNA and made it precipitate. The DNA extracted from the strawberry showed how easy it was to extract DNA out of polyploidy organisms. So if we were to extract DNA out of a haploid organism, it would be very hard to get a big yield of DNA. The result on the mount slide could have varied if the NaCl was contaminated and thus not getting rid of the proteins effectively. This would have resulted in not only DNA on the mount slide but also a great deal of protein. Conclusion: By doing this lab we were able to confirm that DNA from commercial strawberries can be extracted. But we were not sure if the examined DNA under the microscope was exactly DNA. However, the white precipitate formed in the test tube was almost for sure DNA. Also the large amount of DNA being extracted from a single strawberry showed that it was polyploidy and so that polyploidy organisms have a great amount of DNA. This experiment showed the possibility of extracting DNA from commercially available strawberries and examining it.

Sources of Error and Experimental Design Improvements: A possible source of error could be that the white precipitate formed near the top of the test tube wasnt actually pure DNA but rather a mixture of DNA and other molecules. Also since it was not visible under the microscope, it may have been contaminated by dust. As for an experimental design improvement, better filter could have been used to make sure that the correct molecules were being left over. Another possible source of error could have been that the wet mount slide was not prepared properly. Air bubbles may have been left on the slide, distorting the view of the DNA. Post Lab Questions: 1. The purpose of grinding the strawberry was to break down the hard cell wall as well as the cellular and nuclear membranes so that DNA may be extracted. 2. The exact role of the extraction solution is to help us break into the cell nucleus and steal the DNA. This occurs due to the two components of the extraction solution. First the dish soap or detergent helps to dissolve the cell and nuclear membranes. It helps us dissolve the phospholipid biylayer of cell membrane, nuclear membrane and organelles. The membranes are made of lipids (fat) and the detergent will cut through the membrane just like it cuts through grease on a dirty plate when washing dishes. The salt, in this case NaCl, helps separate the proteins from the DNA. It helps us get close to 100% DNA and not the histones (or proteins) that DNA is wrapped around. 3. The main proteins involved when we are talking about DNA are histones. DNA are wrapped around these histones to reduce the space taken up inside the cell. The sodium and chloride ions are more charged and much more polar than the proteins, therefore they displace the histones. Thus the proteins are displaced out of the DNA and so we are left with almost pure DNA. 4. DNA is soluble in water but not in alcohol. In fact, alcohol makes DNA clump together and precipitate out making it visible to the naked eye. Any DNA that contacts the alcohol will clump together, pulling the rest of the DNA strand along behind it. 5. Strawberries, which were used in this lab, are an octoploid meaning they have eight sets of chromosomes. This would also classify them under being polyploidy. This aspect is what makes strawberries a good candidate for demonstrating DNA extraction. Since there are eight copies of each gene in the strawberry genome, they are packed full of it. If we were to use peas for example, which are diploids (contain two sets of chromosomes) then the amount of DNA yielded at the end of the lab would not be as much nor as visible as that with strawberries. Likewise, if we were to use a plant that was haploid (contain a single set of unpaired chromosomes) then the percent yield of DNA for our lab would be much lower compared with strawberries which are polyploidy.

6. No the DNA extracted is not pure. RNA can also be present in the filtrate and some proteins. This is because the filter removes the larger particles from the solution, such as seeds, pith, etc., allowing only the smaller cell components such as the DNA, proteins, etc. to filter through. 7. The genomes of morphologically dissimilar organisms come in a wide variety of sizes. It has been concluded that genome size does not correlated with the complexity of the organism. For example, the genomes of some singled-celled protists are much larger than that of humans. This was explained with the discovery of non-coding DNA. Although some simple organisms contain large genomes, they include non-coding and repetitive DNA which contributes to the size. Whatever the case, there is no clear cut relationship between genome size and organism type. 8. The induction of polyploidy is a common technique to overcome the sterility of a hybrid species during plant breeding. Also polyploidy plants affect crop yield because they make plants such as, apples and strawberries, larger than they are. Polyploidy organisms can photosynthesize faster than haploids and diploids, which create a larger crop yield. There are two ways they are formed. One way is somatic chromosome duplication. This is when chromosomes are duplicated but they are not separated into two cells during meiosis. The amount of chromosomes is then doubled often resulting in a polyploidy. Another way they form is by treating cells with chemicals which inhibit cell division.