Comparison of Methods used for DNA Quantification Method Ease Cost Sensitivity Result UV Spectrophotometry +++++ + ++ Total DNA Yield Gel electrophoresis +++ + + Int vs deg DNA Slot Blot ++ ++ +++ Human DNA Yield Gel blot + ++ +++ Int. vs. deg human DNA Pico-green microtitre plate ++++ ++ ++++ Total DNA Alu Quant +++ +++ ++++ Human DNA Real time PCR assays +++ +++ +++++ Human DNA Real time PCR assays +++ +++ +++++ Int.vs. deg.Human DNA

Summary 1 (from last week) Why study DNA Law enforcement, evolution, agricultural, and human applications-medical diagnostics DNA Biology and Genetics DNA is contained in cells –the basic unit of life Found in nuclei, mitochondria and chloroplasts Organized in chromosomes. Located at positions called loci and come in different forms or alleles. Homozygous if the same, heterozygous if different Alleles segregate independently and assort randomly when on different chromosomes. Random assortment is desired for forensic DNA loci. DNA Function and Structure DeoxyriboNucleic Acid : blueprints of life Replication, Information storage and mutation RIM Central Dogma DNA------->RNA------>protein transcription translation

Summary 2 DNA Structure and Function continued: DNA Replication Bases are Adenine, Guanine, Cytosine and Thymine- Asian Guys Can Teach: AGCT Base pairing is A to T and G to C- DNA is where its AT Sequence of Bases Store information- Like the sequence of numbers in a Phone Number Sides of the ladder are Sugar-Phosphate backbones Nucleotides are the building blocks (dNTPs) themselves made of phosphate base and sugar= PBS- The only station Sierra and Gabriel can watch DNA base pairs- DNA velcro (David Letterman DNA Replication Semi-conservative- Half republican (old) /half democrat (new) Template directed with base pairing (AT, GC) 5 required ingredients of PCR - primer, template, Mg, dntps, DNA polymerase (PTMDD)

Summary 3 Steps in forensic DNA typing are – Evaluation, Extraction, Quantification, Typing and Interpretation 1) Evaluation- Is it there? Screen- blood? Semen? Saliva, human? 2) Extraction- Get and clean DNA Open cells -Get DNA Organic, Chelex and FTA extractions Quantify- Determine quality and quantity? How good and how much did you get? Yield Gels, Slot Blots, Real time PCR assays

Case #2: Can you extract and type DNA from fingerprint powder that has been used to develop latent prints? If so, design an experiment with + and – controls to support your hypothesis.