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1. After re-do your culture, you now have to amplify your report gene so that you...

1. After re-do your culture, you now have to amplify your report gene so that you have enough DNA to continue your experiment. You decided to alter the concentration of template DNA strand as a testing condition. After PCR, you decided to run a gel electrophoresis to see the results of your testing condition. You loaded your ladder and enough DNA to give you bands that are bright enough to visualize. However, after you run the gel and take it under the UV light, you see no bands in samples lands and the ladder lane. What happened?

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Answer #1

Ans. DNA does not fluoresces under UV light on its own. To observe the bands under UV light, the DNA must be chelated with ethidium bromide or other suitable dye like gelGreen.

# Since both the ladder and sample bands are NOT visible under UV light, the following errors might have occurred-

            #I. It is most likely that EtBr treatment has not been done prior to visualization.     #II. The UV lamp might not be working well. For example, EtBr fluoresces at 254 nm, if the UV lamp does not emits light of this wavelength, the fluorescence does not occur, and DNA bands won’t be observed.

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