Extraction And Visualisation Of The Kiwi Fruit And Strawberry Fruit Dna

This paper explores the history and some interesting facts about DNA. The last couple centuries have seen an exponential growth in our knowledge of DNA. The history of the DNA can be traced back to multiple devoted scientist. This article attempts to summarize, and review the basic history of DNA while providing some fascinating information about it.

In this experiment we were meant to observe the transferring of DNA. There are many ways in which DNA can be transferred into an organism, for example; transformation, transduction, and conjugation. In our experiment we used

Restriction Enzyme Digestion – The experiment was begun after putting on gloves to avoid any chemical contact with the skin. Four microtest tubes were obtained, and each of them was labeled to contain the different enzymes or suspect DNA. Two of the microtest tubes were used for suspect one and the two different restriction enzymes, while two other microtest tubes were labeled for suspect two and the two restriction enzymes. After labeling the tubes, the contents that were at the bottom were taken out by slightly tapping them. Then to begin setting up the enzyme reactions, a micropipette was used to obtain 10 μL of the reaction buffer which was added to each of the four test tubes. The buffer is important because it carries the electrical current from the power supply in the gel. After the reaction buffer was in each, the microtest tubes were individually filled with their specific enzymes and DNA, shown in summary through Table 1.1 below. The restriction enzymes are used to cleave the DNA at specific

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.

Four microcentrifuge tubes were placed in a rack, labeled and numbered, in order to identify the group and the DNA/restriction enzyme that it held. Each of the tubes initially received 10 microliters of reaction buffer. There were two samples of suspect DNA provided along with two restriction enzymes (EcoRI and HindIII). Tubes labeled 1 and 2 received 15 μL of DNA from suspect one while tubes 3 and 4 received 15 μL of DNA from suspect two. Following that, 15 μL of Enzyme 1 (EcoRI) were added to tubes 1 and 3, and 15 μL of Enzyme 2 (HindIII) were added to tubes 2 and 4. (Table 1). The tubes were then gently tapped on the counter to mix the DNA and enzyme solution followed by incubation at 37°C for 45 minutes. After incubation, 5 μL of 10x gel loading dye were added to each of the four tubes of suspect DNA. The tubes were then placed on ice while the gel was under preparation.

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).

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

All the procedures performed were according to the Biochemical Techniques: Recombinant DNA Laboratory Second Edition (1). Restriction Digestion of Unknown Vector An unknown vector (0.1 µg/µL) was assigned to the group for restriction enzyme digestions. The vector could be either pUWL 500 or pUWL 501.Three enzymes, Pst I, Bgl II, and Eco RV, and their combinations were used to single, double and triple digest the unknown vector. A total of nine tubes of digestion mixtures were made for this experiment.

The second method was to purify plasmid DNA from the E. coli culture and physcically analyze the DNA by digesting the plasmid with specific restriction endonucleases. The restriction endocucleases used were Bam HI (B), EcoRV (E), and BgIII (G). These restriction endonucleases recognize specific short DNA squences. These cut plasmids were then placed in a 1% aragrose gel to determine the band sizes. The expected phenotype for Pglo is that there will be floursenscens light produced on the LB + amp + Arab plate, where as for PFG the expected phenotype is that there will be flourcent phenotype only in LB+ Amp + IPIV. For the physical analysis Pglo will produce fragment sizes in restriction enzyme (E) 5371bp long. Enzyme (B). fragment sized 625bp, 220bp, and 4,526bp in restriction enzyme (B). Pfg will produce 2 fragments in restriction enzyme (E), one will be around 3,855bp long, and the other one will be 2,506bp long. In restriction enzyme (B) there will be only one band about 6361bp long. From restriction enzyme (G) there should be one bad around 6361bp

The purpose of the experiment was to isolate plasmid DNA, followed by restriction digestion using restriction endonucleases and then visualizing the digested fragments after subjecting to gel electrophoresis. Plasmid DNA (pSP72 DNA) was isolated from Escherichia coli KAM32 (E.coli) cultures using the QIA prep miniprep kit and then subjected to restriction digestion by EcoRI and HindIII. The restriction digested DNA was then loaded into the wells of 0.7% agarose gel and subjected to electrophoresis. It can be concluded from our results that our plasmid DNA isolation was successful and the restriction digestion results were partially in agreement with our hypothesis.

Restriction enzymes cut DNA at certain sites to create multiple DNA fragments. Restriction enzyme HindIII has known DNA fragment lengths and recognition sites when digesting lambda DNA, while the lambda DNA recognition site for restriction enzyme XhoI is unknown. The goal of this study is to determine the lambda recognition site of XhoI by comparing a HindIII digest and a HindIII and XhoI double digest on an electrophoresis gel. The HindIII digest had a band at 9.4 kb, but this band was not visible in the double digest, therefore we concluded the recognition site for XhoI was around 9.4kb. There were also two additional DNA

Analysis of DNA from practicals 1 and 2 using the technique of agarose gel electrophoresis and analysis of transfomed E. coli from practical 2 (part B)

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

Plasmid DNA with Restriction Digest: The purpose of restriction digest of plasmid DNA is to understand how each DNA plasmids was cut with the given restriction enzymes and perform gel electrophoresis to observe the samples. Nine restriction digests were created, containing three digests for each of the three plasmid DNAs identifying as recombinant, non-recombinant, and unknown. Out of the nine digests, six are actual digests and three are undigested controls. A master mix is created to add to each of the nine samples with its following stock ingredients: 10 ul of 2X Reaction Buffer, 1 ul of Nco1, X ul of sterile water (Single digest), 10 ul of 2X Reaction Buffer, 10 ul plasmid DNA, 1 ul Nco1, 1 ul of Not1, and X ul of sterile water (Double