A CCD-based system for the detection of DNA in electrophoresis gels by UV absorption

A method and apparatus for the detection and quantification of large fragme nts of unlabelled nucleic acids in agarose gels is presented. The technique is based on ultraviolet (UV) absorption by nucleotides. A deuterium source illuminates individual sample lanes of an electrophoresis gel via an array of optical fibres. As DNA bands pass through the illuminated region of the gel the amount of UV light transmitted is reduced because of absorption by the DNA. During electrophoresis the regions of DNA are detected on-line us ing a UV-sensitive charge coupled device (CCD). As the absorption coefficie nt is proportional to the mass of DNA the technique is inherently quantitat ive. The mass of DNA in a region of the gel is approximately proportional t o the integrated signal in the corresponding section of the CCD image. This system currently has a detection limit of less than 1.25 ng compared with 2-10 ng for the most popular conventional technique, ethidium bromide (EtBr ) staining. In addition the DNA sample remains in its native state. The rem oval of the carcinogenic dye from the detection procedure greatly reduces a ssociated biological hazards.