Testing of Intercellular Material for DNA through Agarose Electrophoresis Purpose: The point of this lab was to determine whether or not DNA was actually extracted in the prior week’s experiment, in which E. Coli bacteria’s was lysed and through a series of chemical extractions it’s inner contents were harvested. Methods: 4.5 mL E.Coli EDTA suspension was pipetted into a conical tube. After this 0,25 mL lysosome solution was put inside the same tube. Both were incubated at 37°C for so minutes. Once out of the incubator, 0.5 mL of 10% SDS was added. In order to ensure a good mixing of the liquids the tube was inverted continuously for several minutes. Incubation occurred once more but this time at 60°C for 10 minutes. Once cold down to room It was loaded into the gel instead of in the well leading to no results. There should have been a series of bands similar to well three. In lanes three, 5, and 6 we see the bands labeled A,B and C. Due to the short distance traveled down can discern that these bands are in fact DNA as it is both bulky and negatively charged. In lane seven one can see the band in the beginning, which is identified, as tRNA. E and F represent the left of “junk” from the inside of the cell that made it’s way into the sample such as mRNA and proteins. This can be told by seeing that the small materials traveled very far down the gel and were not removed by DNase. What truly tells one whether or not he or she extracted DNA are the blank spots on the gel. In lane four and eight there are missing bands. This is due to the fact in these samples enzymes where added to break down the nucleic acids, DNase in the case of lane four and RNase in eight, thus causing a gap where they should appear. The data that was collected seems to indicate that the sample that was extracted was done properly and yielded DNA. It should be noted that the lanes three and four were switched when adding the material into
Digestion of the haemolytic and non-haemolytic cells allowed for easier identification of fragments during electrophoresis analysis. Lane 12 in figure 3 show the size markers of SPP1 digested with EcoR1 while lanes 6 and 7 show samples of pK184hlyA and pBluescript digested with EcoR1 and Pst1. Lane 4 was loaded with plasmid DNA from haemolytic cells digested with EcoR1 and Pst1 while lane 5 was loaded with EcoR1 and Pst1 digested DNA from non-haemolytic cells. There was a lack of technical success in both lanes due to no bands appearing in lane 4 and only a single band appearing in lane 5. Theoretically, two bands should appear in both lanes after successful to allow for fragment identification. A possible explanation for the single, large fragment in lane 5 is that successful digestion did not take place and the plasmid was only cut at one restriction site leaving a large linear fragment of plasmid DNA. The absence of bands in lane 4 could be because there was not enough plasmid loaded into the lane. Another possibility could be that low plasmid yield as obtained when eluting the experimental samples in order to purify it. Lanes 8 and 9 belonged to another group and show technical success as two bands were present in both the haemolytic (lane 8) and non-haemolytic (lane 9) lanes. If the
Living organisms undergo chemical reactions with the help of unique proteins known as enzymes. Enzymes significantly assist in these processes by accelerating the rate of reaction in order to maintain life in the organism. Without enzymes, an organism would not be able to survive as long, because its chemical reactions would be too slow to prolong life. The properties and functions of enzymes during chemical reactions can help analyze the activity of the specific enzyme catalase, which can be found in bovine liver and yeast. Our hypothesis regarding enzyme activity is that the aspects of biology and environmental factors contribute to the different enzyme activities between bovine liver and yeast.
The given DNA ladder sample and each individual ligation samples were run on 40ml of 0.8% agarose in 1x TAE buffer for approximately sixty minutes at 110V. The appropriate volume of 6x GelRed track dye was used after it was diluted to a final concentration of 1x and incubated for thirty minutes. Finally, the gel was illuminated under UV light and analyzed.
I used a piece of gauze, folded it in half, placed it in a funnel, and used it as a filter. The mixture was filtered in a beaker. Then used 20 mL of ice cold alcohol to pour into the beaker. The alcohol was used to separate the DNA and made it more visible. Lastly, I collected the DNA with a swirling glass stirring rod, patted the DNA dry with a paper towel, and measured the
Then, using a fresh tip each sample, I transferred of the enzyme to each separate tube of the DNA samples. By adding the enzymes, this will cut the DNA molecules into small pieces when we place it into the gel and let it
Paabo’s team, from Leipzig, Germany, used a method of amino acid content as a way of measuring extractible DNA from the bones. The amino acid method was a...
The bands, all except lane 6, which was DNA from Suspect 1 cut with enzyme 1, were visibly shown as lane 6 did not have any bands that were visible to be compared against for crime scenes one or two.
...rm (1:1) was added to the linearized sample in a 1.5 ml microcentrifuged tube. The mixtures were centrifuged at 13000 rpm for 2 minutes at room temperature. The upper aqueous solution was transferred to another sterile 1.5 ml microcentrifuged tube. Equal volume of chloroform was added and centrifuged at 13000 rpm for 5 minutes. Again, the aqueous solution was transferred to a new 1.5 ml microcentrifuged tube and 1/10 volume of 3M Sodium Acetate Solution was added. Then, 2.5 volumes of cold absoluted ethanol were added to precipitate the DNA. The mixture was incubated in -20 °C for overnight or -80 °C for 1-2 hour.
The liquids used were distilled water, and a starch solution, in line with the guiding question. Dialysis tubing was used because it performed similarly to an actual cell while being visible to the naked eye. First, I soaked four strips of dialysis tubing in water for 5 minutes, afterwards I knotted off one of the ends for all of the tubing. After, I filled two with a half tablespoon of distilled water, and another two with half a tablespoon of the starch solution. Next I measured the lengths, widths, and weights of the cells. I proceeded to leave the four cells to soak in distilled water for about 24 hours. After leaving them in distilled water for about a day, I extracted the cells from the distilled water and remeasured the measurements mentioned before. In hindsight, this was an ideal method to investigate the guiding question because the dialysis tubing functions almost identically to a living cell while being easier to observe and handle at the
There are other ways to extract DNA. You can even do it at home with in house items. You can even do this easy experiment in three easy steps. Blend a bag of peas then strain them into a blender. After pouring all that into a cup then poor liquid detergent into the
== § Test tubes X 11 § 0.10 molar dm -3 Copper (II) Sulphate solution § distilled water § egg albumen from 3 eggs. § Syringe X 12 § colorimeter § tripod § 100ml beaker § Bunsen burner § test tube holder § safety glasses § gloves § test tube pen § test tube method = == = =
LAB REPORT 1st Experiment done in class Introduction: Agarose gel electrophoresis separates molecules by their size, shape, and charge. Biomolecules such as DNA, RNA and proteins, are some examples. Buffered samples such as glycerol and glucose are loaded into a gel. An electrical current is placed across the gel.
Then the gel was placed on a white background to make it easier to see the wells. Then using the pipette man and pipette tips, 20 L of sample was added to each well. To avoid contamination, a new tip for every DNA sample is used. Which samples belonged to each well was written down. Once this was done, two carbon fiber electrodes were placed into the gel, one at each end.
doubt be linked to the scene. All that is needed to extract DNA is one
Biology is the study of living organisms divided into specialized fields that cover their morphology, physiology, anatomy, behavior, origin, and distribution. One of the most important fields within biology is microbiology, a field that details the function and behavior of microorganisms that remain invisible to the human eye. Using devices like electron microscopes scientist are able to identify, characterize, and record the morphologies and behaviors of various microorganisms. One of the most essential components of all organisms including microorganisms is their genetic information. With the development of microbiology over the past century, DNA has been identified as the macromolecule that carries genetic information. Some key experimentations