Photosynthesis Lab

After it was expelled back into the cup, 1 ml of the saline rise was transferred into a micro test tube in order for it to be spun in a balanced centrifuge. The micro test tube that contained the 1 ml of saline rise was inside of the centrifuge for 2 minutes, resulting in cells at the bottom of the tube. Since all of the cells were present at the bottom of the tube, the saline was poured off and the tube was vortexed to be sure that there were no clumps of cells. Afterward, the InstaGene Master Mix (which removes cofactor to inhibit DNA cutting enzymes) was vortexed with the saline rise in order to fully mix the contents of the tube (Bio-Rad Laboratories, Chromosome 16: PV92 PCR). The tube was then incubated at 56 degrees Celsius for 10 minutes to inactivate DNAses and put on a second heat block at 100 degrees Celsius for 5 minutes to disrupt cell membranes. The tubes were put into a centrifuge then cooled down in a 4 degrees’ Celsius fridge.

Using the electric current, scientists pass the DNA through gel, and as smaller molecules get through gel quicker than those of bigger sizes, DNA molecules get separated according the sizes of the molecules. We utilize the property that large molecules move slower, and DNA is slightly negatively charged(due to phosphate groups), so it will move to the positive pole of the gel.

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.

On October 12 2017, the class was set to collect DNA samples from a strawberry. Placing the strawberry in a ziplock bag and zip up the bag. Squish the strawberry into a liquid. Add 10 ml of the buffer (salty, soapy water) into a 10ml graduated cylinder using a transfer pipettes. Using the 10ml graduated cylinder, open up the ziplock bag and pour the 10 ml buffer with the squished up strawberry then zip up the ziplock bag. Mix it by moving it around using your fingers, but be cautious about how much bubbles are created. Add the a funnel lined with a cheesecloth onto a test tube (filtration apparatus) and filter the new berry-buffer solution in the filtering apparatus into a test tube. When finished with adding the new berry-buffer solution

: During each reaction, something happened. During reaction 1, after adding the nitric acid a red-brown gas was created. The copper first turned green after a few minutes turned into a blue solution. Reaction 2, When acid was added a blue precipitate formed. Reaction 3, when heated the solution turned into a black precipitate.

≤ 0.05, + Injured berries inoculated with the living bacterial cells at 1x108 CFU mL-1 followed by challenge with different Aspergillus sp. at1x104 CFU mL-1 after 72 h and kept at 20 oC, ++ Intact berries suspended in the suspension of living bacterial cells at 1x108 CFU mL-1, injured and challenged with different Aspergillus sp. at1x104 spores/ mL-1 after 72 h and kept at 20 oC, +++ Intact berries suspended in the crude cell extract from freeze-dried and thawed bacterial cell suspensions at 1x108 CFU mL-1, injured and challenged with different Aspergillus sp. at1x104 CFU mL-1 after 72 h and kept at 20 oC.

Purpose: In doing this lab, the objective was to find how the intensity of light effects the of the products of photosynthesis. This lab will measure to what extent production of photosynthesis increases when light intensity is increased. This experiment will not only provide a visual representation of the process, but will also allow for further examination of the relationship between photosynthesis and cellular respiration. Introduction: The process of photosynthesis uses energy from the sun to produce glucose for plants, and cellular respiration takes glucose and oxygen to form carbon dioxide and water.

Experiment 1 & 2: The DNA concentration was 60 ng/µL (6.00*10^4 ng/mL). The total yield was 0.6 ng (60 ng/µL DNA /100 µL H2O). The DNA sample was divided by 100 µL instead of 25 µL of H2O because the sample was diluted 4 times.

During this lab, spectroscopy and chromatography was used to determine the various properties and characteristics of fast green solution, chloroplasts and an unknown solution. The spectrometer helped determine the absorbance levels of each substance which was used in this experiment. The levels which were determined were used to find the concentration curve of the concentrated solution of fast green solution and the concentration of the given unknown 215. The chlorophyll solution presented a varied distribution in the absorbance levels which would eventually help us determine what wavelengths are absorbed by chlorophyll. Chromatography was used to separate the components of the chlorophyll (spinach) solution which revealed

The main purpose of the lab was to be able to isolate the DNA in the citrus plants and amplify it to determine if it contained the circadian clock gene. The first step that we did in the DNA isolation experiment was receive a citrus leaf and grind it into a fine powder by adding liquid hydrogen to it. We then added extraction buffer, which break down the cell wall, and transferred the 100 ul of the liquid into a microfuge tube. Followed by the addition of 120 ul of 10% SDS, which denatures proteins, then headed the tube. The heating of the tube will help break the hydrogen bonds that hold the two DNA strands together. The microfuge was then placed in an ice bath, which allows the replication and rebuilding of the DNA strands. By centrifuging the microfuge and pipetting it through a miracloth it allows only the liquid that contains the DNA to be extracted. Isopropanol was then added to the now filtered liquid, once spun a minuscule pellet will be seen at the bottom of the tube, which is the DNA. The excess liquid surrounding the DNA pellet will then be extracted and 70% ethanol will then be added which is used to remove the excess buffer that may have still been left in the DNA. The ethanol was then removed and then we added 100ul of resuspension buffer,

Gel electrophoresis is a commonly used laboratory technique employed in biochemistry and molecular biology (ARBL, 2000). The two most conventional types of gels used for DNA electrophoresis are agarose and polyacrylamide (PA). The two substances differ in factors such as resolving power and in the difficulty of setting up and handling them (ARBL, 2000). In comparison to the polyacrylamide, agarose gels are used more commonly as it may also be refrigerated and re-used, and runs horizontally (Reina, 2014). Gel electrophoresis through an agarose channel is used to identify, quantify and purify nucleic acid components (Life Technologies, 2015). The samples of DNA are loaded into wells of agarose gel which is then subjected to an electric current,

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

b). The strawberry’s yield with more DNA than other fruit and it was octoploid, which means the strawberries have eight copies of each type of DNA chromosome. These circumstances make the DNA of strawberry easier to extract and see.

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 Red #40 was mixed with 10 mL of DI water, the solvent. The volume of stock solution needed to prepare these solutions are shown in the second column of Table 3. After obtaining the appropriate micropipettor, 200 uL, it was used to measure the aliquot of the stock solution. This solution was transferred into a volumetric flask and diluted with the 10 mL of DI water. DI water was used as the blank and the solutions were then placed inside the cuvettes which were then placed into the Vernier pH spec-trophotometer. The absorbance and λmax of each solution were then recorded as seen in the third column of Table 2. These values were then plotted on a linear regression as seen in Figure 1. Next, 0.50 mL of maraschino cherry juice was obtained by using the appropriate micropipet-tor, 100 uL, of the aliquot of the stock solution. This was transferred into a volumetric flask and diluted with nanopure water until the solution reached 25 mL. Nanopure water was used as the blank and the solution was then placed in a cuvette which was then measured inside the Vernier pH spectrophotometer. The absorbance and λmax of this solution was then recorded in third column of Table 3. The cherry juice and nanopure water solution was calculated by using the linear regression equation seen in Figure 1. The cherry juice concentration and nanopure was 2.26 uM. The stock solution was calculated by using m1v1=m2v2. The cherry juice concentra-tion and nanopure water mass, uM, was m1 and the 25 mL of the diluted solution was v1. The v2 was the 0.50 mL of the cherry juice. After calculations, the m2 was calculated. It ended up being 131 µM which was the mass of the stock