RNA concentration is measured to analyze genetic material for diagnostic, research, and forensic purposes. Technicians extract RNA from a sample, process it to limit contamination, and measure it using a spectrophotometer or dye. Low RNA concentration can lead to errors, requiring a new sample or purification.
The concentration of ribonucleic acid (RNA) is a measure of how much of this genetic material is present in a sample. This nucleic acid is one of the important building blocks of life, essential for the functioning of organisms, from whales to domestic cats. It can be analyzed in tests for a variety of reasons, including diagnostic purposes, research and forensic analysis. Before it can be tested, it must be carefully processed and checked to confirm that the sample is of good integrity and produces accurate results.
During processing, technicians extract RNA from a sample so that it can be analyzed. This may involve treatment with enzymes that remove deoxyribonucleic acid (DNA) and proteins that may be present in the sample. Careful controls are needed to limit the chances of contamination and preserve as much RNA as possible. Specialized laboratory glassware and plastic can be used for this process, and technicians also follow a standard laboratory procedure for consistency.
The classic approach to measuring RNA concentration involves running a sample through a spectrophotometer, a device that measures the absorption of light. The readings can provide insight into how much RNA is present based on how much light is absorbed at specific wavelengths. This can also indicate whether there are contaminants in a sample, because other materials absorb light in different wavelengths. Thus, the test can serve a dual function by quantifying RNA and evaluating it for contamination.
Another option is to add a dye to the sample and expose it to light to see if the dye fluoresces and how brightly. Dyes can bind tightly to nucleic acids to provide information about their concentrations. A drawback of this method is that RNA concentration values can be erroneous if the sample also contains DNA or other impurities, as the dye can bind to these as well. Technicians can only use this option to determine RNA concentration if they are sure the sample is very pure to avoid false results.
If the RNA concentration is too low, the sample may not be usable. For example, it may not be enough to run tests and double-check the results. A higher chance of error can also be a problem, because problems with the sample would be magnified if only a limited amount of RNA is available. The technician may need to purify a new lot or request a new sample, if that is an option, to determine if a cleaner sample with a higher RNA concentration can be obtained.
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