wide range of concentrations, we can use 50 µl in a second assay tube. Now the sample
can be as concentrated as 40 mg/ml and we will still have 4 mg or less in the assay tube,
giving a readable result. To cover all of the bases we can assay a third tube with just 5
µl sample
It helps to have a reasonable estimate of ranges of concentrations of sample that one
can expect. Even with such an estimate it is good to prepare samples with a range of
dilutions, in case a sample is so concentrated that its absorbance readings are out of
range.
For the assay in the example, if we use 500 µl sample in an assay tube (the maximum
volume), its concentration would have to be less than 4 mg/ml to give a readable
absorbance. On the other hand, we would want that much if the sample was, say, ten
times less concentrated. Knowing nothing about the concentration of a particular
sample, we would load one tube with 500 µl to cover that range. Since the assay spans a
wide range of concentrations, we can use 50 µl in a second assay tube. Now the samplecan be as concentrated as 40 mg/ml and we will still have 4 mg or less in the assay tube,giving a readable result. To cover all of the bases we can assay a third tube with just 5µl sampleIt helps to have a reasonable estimate of ranges of concentrations of sample that onecan expect. Even with such an estimate it is good to prepare samples with a range ofdilutions, in case a sample is so concentrated that its absorbance readings are out ofrange.For the assay in the example, if we use 500 µl sample in an assay tube (the maximumvolume), its concentration would have to be less than 4 mg/ml to give a readableabsorbance. On the other hand, we would want that much if the sample was, say, tentimes less concentrated. Knowing nothing about the concentration of a particularsample, we would load one tube with 500 µl to cover that range. Since the assay spans a
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