Extraction of DNA from onions

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.

The mixture was cooled down to room temperature and transferred to a separatory funnel where it was washed with four portions of 50 mL of water.

Introduction: All organisms have DNA (Deoxyribonucleic Acid) in the nuclei of their cells. The aim of this experiment is to extract this DNA using common household detergent to breakdown the cellular membrane then through the use of ethanol to extract the DNA from the Kiwi and Strawberry fruit. After the DNA has been extracted the DNA will be visualised by agarose gel electrophoresis which is done by adding a dye to the extracted DNA then being placed in the gel, once the negatively charged DNA has been placed in the gel it will slowly move towards the positively charged end (anode) then is viewed under an ultraviolet light then compared to the control of known lengths of DNA to show the size of the unknown DNA samples.

Deoxyribonucleic acid, DNA for short, is a substance in all living organism that duplicates itself. It is the carrier of all genetic information, such as eye color, hair color, height, etc. DNA is found in all living things including both eukaryotic and prokaryotic organisms. DNA is tightly packaged in the chromosomes to start DNA replication. During this process, DNA unwinds so that it can be duplicated. DNA can be extracted from strawberries which can also explain how DNA can be used for medicinal purposes as well as explaining how DNA extraction originated.

There are three specific steps required to isolate DNA from its cellular contents. The steps used to remove and expose DNA from its cell are: breaking down the food type you are using by crushing it, for example a banana or strawberries, exposing the substance to a sodium chloride (NaCl) solution, subjecting the product to detergent solution (dH2O), filtering the solution and lastly, the addition of ethanol. When beginning with a solid substance, such as a banana, crushing the substance allows for

I looked at he pGLO through a uv light. Then I put my loop into the pGLO plasmid DNA tube and made sure that the ring was covered with he solution, then mixed it into the +pGLO. I then place the tubes back on ice for 10 minutes and I pushed them under the ice. While my tubes were sitting in the ice I begin Ito label my LB nutrient Agar plates on the bottom. When the 10 minutes is up for the tubes in the ice, I removed them from the ice and put them into a film holder and placed them into a heated water bath for 50 seconds.The settings for the water should be 42 degrees Celsius. When the 50 seconds are up, I placed both tubes back on the ice and had them sit in there for 2 minutes. After the 2 minutes I I removed

charged DNA to bind the positively charged minicolumn. 2 ml of column wash is then used to

Next heat shocks the tubes for 50 seconds, followed by icing for 10 more minutes. The heat shock increases the permeability of the cell membrane to DNA. Then add 250 milliliters of LB and incubate for 20 minutes. The 20 minute incubation following the addition of LB broth allows the cells to grow and express the ampicillin resistance protein, beta-lactamase, so that the transformed cells survive the subsequent ampicillin selection plates. Plate 100 microliters the + tubes evenly on two plates; 1 of LB and Amp, and one of LB, AMP, and ARA. Plate 100 microliters of the – tubes evenly on two plates; 1 of LB and AMP, and one on LB only.

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.

After that, a new sterile loop was used to immerse the pGLO plasmid DNA stock tube into the +pGLO tube but no into the – pGLO tube. Both tubes were well closed and put back on the ice for 10 minutes. While both tubes were sitting on ice, the four plates were labeled. LB/amp plate was labeled +pGLO, LB/amp/ara plates were labeled +pGLO, LB/amp plate was labeled – pGLO, and LB plate was labeled – pGLO. After that, the tubes were transferred into a water bath set at 42° C for exactly 50 seconds, then placed rapidly back on the ice for another two

Let sit at room temperature for 2.5 minutes 9. You will start to see a cloud film develop into the layers of solution. (This is your DNA) 10. Gently spool the DNA onto a glass rod, and dissolve into 1000µ of 1xTE buffer. 11.

Since the temperature optimum for the DNA polymerase to act is 72° C, the reaction is heated to that temperature. DNA polymerase lengthen the primers by attaching more nucleotides onto the primer in a sequential manner, using the target DNA as a template.

To start this laboratory, it must first label two transformation tubes of different color; one with – DNA and the other with + DNA. Then, using a P-1000 micropipette adds 250 ul of the ice cold transformation solution (CaCl2) into each tube and places both tubes on ice. Using a disposable sterile inoculating loop to transfer colonies of E. coli bacteria from the starter plate to the +DNA tube. Immerse the loop in the CaCl2 solution in the transformation tube and spin the loop until all bacteria is incorporated into the transformation solution. Repeat this step with the – DNA tube. Using a P-10 pipette, transfer 10 ul of pGLO directly into the + DNA tube. Tap the tube lightly with a finger to mix. Place both tubes onto ice, and incubate for 10 minutes.

The Material floating at the top of the test-tube after the alcohol was added was all the strands of DNA. The alcohol dissolved the cell walls of the cells and the DNA was free to move about freely.

DNA, Deoxyribonucleic Acid, is the basic structure for all life, it is the blueprint, the instruction manual, on how to build a living organism. DNA is made up of four nitrogen bases, adenine, thymine, cytosine, and guanine which are connected by sugar-phosphate bonds. Through a process called Protein Synthesis, the nitrogen bases are the code for the creation of amino acids. Essentially, DNA makes amino acids, amino acids make proteins, proteins make organisms. This process has been taking place for much longer than scientists have been able to document. Those scientists are called geneticists and their field is genetics.