Conventional PCR using agarose gel electrophoresis detection Essay

Conventional PCR using agarose gel electrophoresis detection - Essay fountWhile addition of gel, the care for the serving of it has to be taken as a 0.7% gel will show good separation (resolution) of large deoxyribonucleic acid fragments (510kb) and a 2% gel will show good resolution for small fragments (0.21kb). So, the percentage of the gel is kept between 0.7% to 2%.With intention to feed away very tiny fragments, addition of high percentage ( up to 3%), is not useful as a vertical polyacrylamide gel is more assume in this case. The medium percentage is always recommended as low percentage gel may break while trying to lift them and high percentage gels may often brittle not setting evenly. Lewis recommends 1% gel to use.While suggesting for gel tank Lewis recommends, Small 8x10cm gels (minigels) are very popular and give good photographs. For the applications of Southern and Northern blotting, larger gels are used. 3050mL and 205 mL of agarose is required for minigel and la rger gel respectively. While deciding the amount of DNA to be added to this solution, the nature of analysis has to be kept in mind. According to Lewis Typically, a band is easily visible if it contains about 20ng of DNA.After doing all the above cooking Lewis says, I usually digest and load 24L of the 50L obtained from a kit miniprep. But you see how it depends on the quash and size of the bands expected. For PCR reactions, it depends on the PCR but in routine applications 1020L should be plenty to see the product on the gel.Depending on the stack of DNA being loaded and the number of samples, the design of weed out is decided to include in the process. Lewis recommends, Combs with many tiny teeth may hold 10L. This is no good if you want to load 20L of restriction digest plus 5L of loading buffer. When deciding whether a comb has enough teeth, remember that you need to load at least one marker lane, preferably two. After