Micro-Volume Purity Assessment of Nucleic Acids using A260/A280 Ratio and Spectral Scanning
نویسنده
چکیده
The spectrophotometric analysis of biomolecules for sample quantification and purity assessment has been well documented. Quantification is made possible by the absorption of ultra-violet light by chromophores contained in nucleic acids and protein. Consistent with these methods is the use of the Beer-Lambert Law that relates physical property of light absorption by molecules to concentration within the sample by the following equation:
منابع مشابه
Nucleic Acid Purity Assessment using A260/A280 Ratios
A common practice in molecular biology is to perform a quick assessment of the purity of nucleic acid samples by determining the ratio of spectrophotometric absorbance of the sample at 260 nm to that of 280 nm. Here we describe the basis of A260/A280 ratio determinations of nucleic acids and demonstrate the utility of the BioTek PowerWaveTM 200 scanning microplate spectrophotometer to perform t...
متن کاملNucleic Acid Purity Assessment using A260/A280 Ratios
A common practice in molecular biology is to perform a quick assessment of the purity of nucleic acid samples by determining the ratio of spectrophotometric absorbance of the sample at 260 nm to that of 280 nm. Here we describe the basis of A260/A280 ratio determinations of nucleic acids and demonstrate the utility of the BioTek PowerWaveTM 200 scanning microplate spectrophotometer to perform t...
متن کاملThe Importance of the 240 nm Absorbance Measurement
The assessment of nucleic acid purity by the A260/A280 ratio has been used for a number of years. This measurement, however, does not provide all the information that investigator needs to assess nucleic acid preparations. The detection of impurities common to nucleic acid preparations that are not detected with absorbance measurements at 260 nm and 280 nm can in most instances be accomplished ...
متن کاملPurification of intact nucleic acid from samples
is required for many molecular biology applications. Accurate PCR replication of RNA and DNA sequences requires complete removal of cellular lipids and proteins. Likewise, most restriction endonucleases used to digest genomic DNA are inactivated or degraded by cellular proteases normally present prior to purification of the nucleic acids. Thus, failure to remove these cellular contaminants ofte...
متن کاملSpectral analysis of the conformation of polyadenosine diphosphoribose. Evidence indicating secondary structure.
Based on boronic acid chromatography, a rapid method was developed for the purification of polyadenosine diphosphoribose oligomers of various chain lengths. Polyadenosine diphosphoribose oligomers obtained by the new method were distinguished on the basis of spectral criteria. The purity of polyadenosine diphosphoribose was determined by high performance liquid chromatography, which is a highly...
متن کامل