Extract Blend
Extract Blend
Why is it necessary to use reagents in the extraction of DNA Cold? How can you confirm something is DNA extracted?
We extracted DNA from Pea with mixture, then mixing the resulting soft things with pineapple juice and alcohol. Some substances such as mucus float. How can I confirm that the DNA? Also, why using cold pineapple juice?
Well, I have extracted DNA from cells E. coli for some time now and use the pineapple juice is new to me. In the procedure I use, called the mini-prep plamid, I first centrifuge to isolate the bacterial cells them from the liquid medium and then re-suspend cell pellet in medium, TE + RNase, which acts as a preservative and inhibits the action of the enzyme. This solution is to get the cells ready to be aligned, so that in the experiment, it seems as if this could be skipped since the Pea mixed and mixing may have a line of bean cells. Our next step is the use of NaOH for remove DNA from the cells by alkaline lysis. After this step, the DNA free in solution. Since the experiment you did not do any of these steps, the mixture should have extracted DNA into the solution, thus serving the purpose of breaking open the cells. In my protocol, the next step is the addition of acid to neutralize the basis that we previously added. Function other acid added to a solution as this point is that the acid will trigger the protein and chromosomal DNA from the mixture. I think pineapple juice is acidic, so that the juice is served purpose of precipitating cellular DNA and proteins from all the "garbage" that is contained in the mixture. Furthermore, in my protocol, and you, alcohol is used. Alcohol use for the sole purpose of precipitating the DNA. All reagents used in the lab I have to stay cold all the time ... we always start the experiment by getting a bucket of ice, and all reagents are kept ice until ready for use. DNA, once extraced, always stored in a freezer -20 degrees Celsius. This is caused by DNA instabilty. There are many ways for DNA to change denaturalize, and if the reaction conditions even slightly off, even by a few degrees, the DNA will denature and really useful for experimental purposes. There are several way for you to prove this is DNA. You can run a sample of your solution on agarose gel, which is a special tool used to separate strands of DNA and determine DNA length in base-pairs. if DNA, the DNA bands characteristic will appear after the gel has been run and photographed. After running the samples on the gel, the next step will be sequenced. Sequencing quite expensive, so the presence of DNA must be verified before perfoming this task. I work at a molecular biology laboratory, I do these kinds of experiments all the time ... i plasmid DNA isolation yesterday E. coli, for example. So, if you have more questions about this I would be more than happy to try to answer it for you!
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