First improvements in the detection and quantification of label-free nucleic acids by laser-induced breakdown spectroscopy:Application to the deoxyribonucleic acid micro-array technology

摘要

The accurate quantification of nucleic acids is essential in many fields of modern biology and industry,and in some cases requires the use of fluorescence labeling.Yet,in addition to standardization problems and quantification reproducibility,labeling can modify the physicochemical properties of molecules or affect their stability.To address these limitations,we have developed a novel method to detect and quantify label-free nucleic acids.This method is based on stoichiometric proportioning of phosphorus in the nucleic acid skeleton,using laser-induced breakdown spectroscopy,and a specific statistical analysis,which indicates the error probability for each measurement.The results obtained appear to be quantitative,with a limit of detection of 10~5 nucleotides/mu m~2 (i.e.2X10~(13) phosphorus atoms/cm~2).Initial micro-array analysis has given very encouraging results,which point to new ways of quantifying hybridized nucleic acids.This is essential when comparing molecules of different sequences,which is presently very difficult with fluorescence labeling.