40. Do you make 50 mls of 0.8% agarose to run DNA? (4 pts) 41. You have just com

Question

40. Do you make 50 mls of 0.8% agarose to run DNA? (4 pts) 41. You have just completed your plasmid prep. The Nanophotometer indicates the concentration of DNA is 2635 ng/ul. Now you have to load 20 ng of DNA into the well of a gel that holds a maximum of 200 ul. Describe how you would accomplish this considering the limitations of the equipment available in this lab? (20 pts) 42. You have a stock solution containing 6300 ng/ul of DNA. You want to to load 500 ng of DNA into a well that can hold 40 . Describe how you would accomplish this considering the limitations of the equipment available in this lab? (20 pts)

Explanation / Answer

1. Yes, 0.8% 50ml agarose gel can be used for DNA electrophoresis. The gels that are used for Agarose gel electrophoresis or DNA gel electrophoresis normally range from 0.7% to 2% in strength. The strength of the agarose gel used in a particular study is determined depending on the length of the DNA strands or fragments that are to be run on that particular gel. For smaller fragments, a denser gel with a strength of 2% provides good resolution. For larger fragments, less density of the electrophoresis gel is preferred. If the DNA fragments under experimentation belong to the size range of 5-10 kb, then an agarose gel of strength 0.8% would show sufficiently good separation. However, for smaller fragments (like 0.2-1 kb), a 0.8% gel would fail to show proper separation after run-time is over. Speaking of the volume of agarose gel, 50ml is sufficient for carrying out electrophoresis in the smaller gel rigs (gel electrophoresis apparatus).

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