Evaluation of fluorescence based quantification of DNA: Influence of an external fluorescent probe

Much advancement in biological studies has led to the greater understanding of pathological conditions, their diagnosis and treatment modalities. One among them is polymerase chain reaction (PCR) an effective and inexpensive method used in molecular biology for deoxyribose nucleic acid (DNA) amplification. DNA quantification and characterisation is an important step in molecular cloning. Common laboratory techniques like absorption spectrophotometry, spectrofluorometry and real time PCR are used to quantify DNA. Spectrofluorometry is more specific and sensitive than spectrophotometry method. Fluorescence spectroscopy can be used by adding probes to the sample, called fluorescence probing technique. In the present work, animal DNA is extracted from liver tissues initially. The isolated DNA is then treated with certain fluorescence probe, such as methylene blue (MB). This probe can intercalate with DNA conformations and also can be used for quantification of DNA. The interaction studies of MB with DNA gives understanding of the nano-environment of the conformations of DNA. Molecular mechanics calculations are carried out to ascertain the findings from the experimental data.