Janie Sigmon1, Fred Chernomas2 and Lyndon L. Larcom2
1Greenville Hospital System/Clemson University Biomedical Cooperative
2Clemson University, Clemson SC
The RFLP method of DNA fingerprinting was modified to provide information about mixed DNA samples. DNA was extracted from fresh human blood samples and digested with Hae III or Hinf I. Different dilutions of the DNA digests and mixtures of the samples were electrophoresed in agarose gels. After Southern blotting of the DNA onto a nylon membrane, biotinylated oligonucleotide probes (CAC)5 were used to mark identifying bands. Chemiluminescent detection of the probes produced distinct banding patterns on X-ray films of the exposed membranes. Techniques of experimental design and data analysis were developed to determine the relative amounts of DNA in a sample. Using these techniques, quantitation of absolute fragment concentrations can be determined more accurately than by the currently established methods.
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