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Question
USING THIS ARTICLE LINK: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652015000200691&lng=en&tlng=en
Answer as many questions as you can, if you can't answer all. I understand.
1- What is the major idea of the paper? Which technique gave the best result? (2 points)
2- How did they get rid of the soil in the samples (before testing the 4 techniques)? Were they able to get rid of all the soil in the samples (again before testing the 4 techniques)? (3 points)
3- What did they use (enzymatic, chemical or physical or all combined) to extract DNA from the samples in EACH procedure. (12 points)
4- When the DNA was extracted some methods did not lead PCR products. Why was that? (3 points)
Explanation / Answer
Please find the answers below'
Answer 1: According to the information, the present investigation was targetted to extract maximum quality and quantity of DNA from nematode samples using four different methods. After carefully conducting all the experiments, it was found that the CTAB-method in combination of use with proteinase K emerged as best method.
Answer 2: The soil was separated from the biological sample by using the technique called centrifugal-flotation method. The researchers were able to separate all the soil from the biological sample using this method and thus further extraction steps were performed on nematodes.
Answer 3: All methods were based upon use of combination of either chemicals or chemicals with enzymes discretely. For example, tris-buffer was primarily used in method 1, SDS mediated degradation of membranes and proteinase K mediated digestion of cellular protein was obtained in method 2, chemical and mechanical break down of sample using commercially available resin was performed in method 3 and finally, CTAB-buffer along with proteinase K mediated digestion was performed in method 4.
Answer 4: Two possible reasons why PCR products were not available in some methods are:
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