Consequences of various contaminants on RNA quantification and quality controls
نویسندگان
چکیده
RNA isolation is performed everyday by molecular biologists for a wide range of applications. No matter what protocol is used and how meticulous the user is, a risk to degrade RNA during the process or to carry over contaminants always exists which may affect the reliability of the subsequent analytical techniques. The parameters that are usually checked for quality control include the concentration, the ratios of absorbance A260/280 and A230/260 on a spectrophotometer and the RNA Integrity Number (RIN) used by the Agilent Bioanalyzer. However, biologists new in the field of RNA are often puzzled when these parameters vary from the expected optimal values. The most common problems are RNA partially degraded and RNA contaminated by protein, genomic DNA, or reagents used during purification. Here we provide examples of Bioanalyzer electropherograms and nanodrop values resulting from various type of contamination or degradation. These can be used for guidance to identify the causes of problems in RNA preps. We envisaged:
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