Why does the calculated amount of RNA in solution differ from that on the Product Transfer Form?
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The sample may not be thoroughly mixed. Upon drying, RNA may form aggregates or higher-order structures. To disrupt these, heat samples to 95°C for 1-3 minutes and slowly cool for 30-45 minutes to reanneal complementary strands.
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Differences in instrumentation used for quantifying RNA may lead to differences in the apparent values observed. Consider using dual-beam UV-VIS spectrophotometers for accurate measurements.
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The sample is too concentrated. Absorbance values are most accurate between 0.15 and 0.6 and within the linear range of a standard curve.
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The sample is too diluted. Measurements with dilutions of small volumes (1-1.5 uL) are more susceptible to variation.
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02/19/2008