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Thermo Scientific NanoDrop SpectrophotometersNucleic AcidBest PracticesSampling TechniqueVolume RequirementUse adequate sample volume to ensure good column formation.Tip: Although 1 μL volumes are usually sufficient for most samplemeasurements, increasing the sample size to 2 μL will ensure propercolumn formation for samples with reduced surface tension.Pipettor SelectionUse calibrated pipettor with well-fitting tips.It is best to use a precision pipettor (0 – 2 μL) with low retention,precision tips to ensure that sufficient sample (1 – 2 μL) is delivered foroptimal column formation.Tip: To avoid evaporation errors, it is essential to use an eight-channelpipettor to simultaneouslyload samples when using two or morepedestal positions on the NanoDrop 8000.Sample AliquotsAlways use fresh tips and fresh aliquots for every measurement.Tip: Repeated measurements on the same sample aliquot will result inevaporation, yielding increasing concentrations and/or column breakage.Sample HomogeneityHighly concentrated nucleic acid samples require careful attention toensure homogeneity before sampling.Tip: Non-reproducible results observed when making small volumemeasurements are a good indicator that the sample is not fully in solutionor is not homogenous.Tip: It may be necessary to heat and/or lightly vortex samples prior tosampling to ensure homogeneity.Sample PreparationEnsure sample isolation procedure is optimized and sample is purifiedprior to measurement.8
Thermo Scientific NanoDrop SpectrophotometersNucleic AcidBest PracticesSample PurificationAny molecule that absorbs at 260 nm will contribute to the totalabsorbance value used to calculate sample concentrations. Examplesinclude DNA, RNA, free nucleotides, proteins and some dyes, as well asmany other molecules found in common buffers.Tip: To minimize the potential of overestimating a nucleic acid sampleconcentration as a result of an interfering substance, it is important topurify samples prior to making absorbance measurements.Lab Notes9
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