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Forensic Biology by Richard Li

Chapter 16: Detection Methods. Forensic Biology by Richard Li. Direct Detection of DNA in Gels. Agarose Gels Ethidium Bromide Amounts as little as 10 ng per band Including in the gel OR staining after electrophoresis UV transilluminator at approx. 300nm & orange filter

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Forensic Biology by Richard Li

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  1. Chapter 16: Detection Methods Forensic Biologyby Richard Li

  2. Direct Detection of DNA in Gels • Agarose Gels • Ethidium Bromide • Amounts as little as 10 ng per band • Including in the gel OR staining after electrophoresis • UV transilluminator at approx. 300nm & orange filter • Denatured Polyacrylmide Gels • Silver Nitrate • 100x more sensitive, lower cost, safer Forensic Biology by Richard Li

  3. Hybridization-Based Assay • Radioisotope Labeled Probes • VNTR testing & DNA quantification • Nick translation- dNTPs • Dnase I • P- most common radioisotope • Enzyme Conjugated Probes with Chemiluminescence Reporting System • Alkaline Phosphatase • Lumigen PPD Forensic Biology by Richard Li

  4. Hybridization-Based Assay • Biotin • Vitamin H found in egg yolk • Enzyme Conjugated Avidin • Glycoprotein found in egg whites • Binds to Biotin • Streptavidin • Horseradish peroxidase- contains Heme Forensic Biology by Richard Li

  5. Hybridization-Based Assay • Reporter Enzyme Assay • TMB- chromogenic substrate • Luminol-chemiluminescent substrate Forensic Biology by Richard Li

  6. Detection Methods for PCR-Based Assays • Fluorescence Labeling • Dyes • 400-600 nm • Labeling Methods • Primer method for STR (one primer) • Fluorescently labeled ddNTPs • Fluorophore Detection • Laser utilized as excitation source • Optical filters used to detect emitted light • Detector Forensic Biology by Richard Li

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