It is now more than ten years since Dr. Alec Jeffreys (now Professor Sir Alec Jeffreys, FRS) reported in Nature that the investigation of certain minisatellite regions in the human genome could produce what he termed DNA fingerprints and provide useful information in the fields of paternity testing and forensic analysis. Since that time we have witnessed a revolution in the field of forensic identification. A total change of technology, from serological or electrophoretic analysis of protein polymorphisms to direct investigation of the underlying DNA polymorphisms has occurred in a short space of time. In addition, the evolution and development of the DNA systems themselves has been rapid and spectacular. In the last decade we have progressed from the multilocus DNA fing- prints, through single locus systems based on the same Southern blot RFLP technology, to a host of systems based on the PCR technique. These include Allele Specific Oligonucleotide (ASO)-primed systems detected by dot blots, the binary genotypes produced by mapping variations within VNTR repeats demonstrated by minisatellite variant repeat (MVR) analysis, and yet other fragment-length polymorphisms in the form of Short Tandem Repeat (STR) loci. Hand in hand with the increasing range of systems available has been the development of new instrumentation to facilitate their analysis and allow us to explore the possibilities of high volume testing in the form of mass scre- ing and offender databases.

Klappentext

In Forensic DNA Profiling Protocols, leading forensic experts from around the world describe in detail their time-proven methods for identity testing through DNA analysis. Their state-of-the-art collection of easily reproducible methods includes all of the major techniques of DNA analysis currently used in forensic identity testing. The book covers PCR-based test systems, the now widely used STR typing systems (complete with details of both manual and automated detection systems), repeat unit mapping (MVR), direct-phase minisequencing, capillary electrophoresis, and direct blotting. Also fully discussed are the recovery of DNA from a wide range of sample types (blood, semen, skeletal remains, and saliva), species testing, sex determination, and mitochondrial DNA testing.

Forensic DNA Profiling Protocols captures and makes practical for all laboratories the whole array of new technologies and techniques that have revolutionized forensic science. The readily reproducible methods elucidated here represent the state-of-the-art for those scientists who want to introduce, establish, and perfect DNA identification techniques in their laboratories.

Inhalt
Recovery of High-Molecular-Weight DNA from Blood and Forensic Specimens.- Recovery of DNA for PCR Amplification from Blood and Forensic Samples Using a Chelating Resin.- PCR Analysis of DNA from Fresh and Decomposed Bodies and Skeletal Remains in Medicolegal Death Investigations.- PCR Analysis from Cigaret Butts, Postage Stamps, Envelope Sealing Flaps, and Other Saliva-Stained Material.- Quantification of DNA By Slot-Blot Analysis.- Rapid Assessment of PCR Product Quality and Quantity by Capillary Electrophoresis.- Southern Blotting of Genomic DNA for DNA Profiling.- Preparation and Use of Alkaline-Phosphatase-Conjugated Oligonucleotide Probes for Single-Locus and Multilocus VNTR Analysis.- Preparation and Use of 32P-Labeled Single-Locus VNTR Probes in Identity Testing.- Statistical Methods Employed in Evaluation of Single-Locus Probe Results in Criminal Identity Cases.- Interpretation and Statistical Evaluation of Multilocus DNA Fingerprints in Paternity and Relationship Testing.- Overview of PCR-Based Systems in Identity Testing.- An Introduction to PCR Primer Design and Optimization of Amplification Reactions.- Analysis of Amplified Fragment-Length Polymorphisms (VNTR/STR Loci) for Human Identity Testing.- Amplification of Short Tandem Repeat Loci Using PCR.- Manual Electrophoretic Methods for Genotyping Amplified STR Loci.- 17 Genotyping STR Loci Using an Automated DNA Sequencer.- Separation of PCR Fragments by Means of Direct Blotting Electrophoresis.- 19 Amplification and Sequencing of Mitochondrial DNA in Forensic Casework.- Minisatellite Variant Repeat Unit Mapping Using PCR (MVR-PCR).- Sex Determination by PCR Analysis of the X-Y Amelogenin Gene.- Species Determination by Analysis of the Cytochrome b Gene.- Use of the AmpliType PM + HLA DQA1 PCR Amplification and Typing Kits for Identity Testing.- The Use of Capillary Electrophoresis in Genotyping STR Loci.- Solid-Phase Minisequencing as a Tool to Detect DNA Polymorphism.