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Quantification made easy

With the aid of spectroscopy, the quantitative analysis of nucleic acids and proteins has established itself as a routine method in many laboratories. It includes absorption measurements in the ultraviolet and in the visibility range. Proteins are measured (directly) at 280 nm, nucleic acids at 260 nm and colorimetric protein determination is carried out in the range from 550 to 600 nm. The BioPhotometer offers the following pre-installed test procedures: 1. Nucleic acid determination

DNA, RNA, oligonucleotides and even mononucleotides can be measured directly in aqueous solutions in a diluted or undiluted form. Aqueous buffers with low ion concentrations (e.g. TE buffer) are ideal for this method. The concentration is determined by measuring at 260 nm against blank and then evaluating via factor. Normally, the user has to calculate the concentration of the measured sample using according factors. In contrast, the BioPhotometer can change these factors easily and will do all necessary calculations.

The absorption of 1 OD (A) is equivalent to approximately 50 g/ml dsDNA, approximately 33 g/ml ssDNA, 40 g/ml RNA or approximately 30 g/ml for oligonucleotides. Purity determination of DNA Interference by contaminants can be recognized by the calculation of ratio. The ratio A260/A280 is used to estimate the purity of nucleic acid, since proteins absorb at 280 nm. Pure DNA should have a ratio of approximately 1.8, whereas pure RNA should give a value of approximately 2.0. Absorption at 230 nm reflects contamination of the sample by substances such as carbohydrates, peptides, phenols or aromatic compounds. In the case of pure samples, the rat
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