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Bio383H Lab 1 discussion p. 2 draft

Submitted by liamharvey on Thu, 02/01/2018 - 19:25

The molecular absorbance spectroscopy via Nanodrop showed very different results. In regard to concentration, DNA1, DNA1 with RNase and DNA2 with RNase all showed concentrations around 150ng/µL. However, DNA2 showed concentration of only 91.3ng/µL. This low concentration could be due to errors when making dilutions or perhaps when using the Nanodrop itself. DNA1 and DNA2 had 260/280 values of 2.07 and 2.04 respectively. 260/280 values over ~1.8 suggest that there is a high concentration of RNA in the sample. For DNA1 with RNase and DNA2 with RNase, values for 260/280 were recorded at 2.95 and 3.45. These results would suggest that the two RNase treatments would have more RNA present than the original samples. This result does not agree with the gel electrophoresis results. In the gel electrophoresis it was clear that the RNase treatments did not have RNA present. Perhaps the presence of RNase interfered with the Nanodrop, or there were issues with the computer. DNA1 and DNA2 had 260/230 values of 1.65 and 1.51. 260/230 values below ~2.0-2.2 suggest that carb carryover occurred. Perhaps not all of the carbohydrates which should have been removed via KAOc were indeed removed. DNA1 with RNase and DNA2 with RNase had 260/230 values of -1.43 and -0.73. These values are under the ~2.0-2.2 range, however, they are negative which could mean something else has occurred. The 260/230 value is a ratio, so a negative value may indicate something other than carb carryover. The negative values could also be another possible error with the Nanodrop.

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