Forensic DNA Profiling ProtocolsPatrick J. Lincoln, Jim Thomson 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. |
Contents
Interpretation and Statistical Evaluation of Multilocus | 97 |
An Introduction to PCR Primer Design and Optimization | 121 |
Analysis of Amplified FragmentLength Polymorphisms VNTRSTR | 155 |
Amplification of Short Tandem Repeat Loci Using | 173 |
Manual Electrophoretic Methods for Genotyping Amplified STR Loci | 181 |
Genotyping STR Loci Using an Automated DNA Sequencer | 193 |
Separation of PCR Fragments by Means of Direct Blotting | 209 |
Preface v | v |
Quantification of DNA by SlotBlot Analysis | 33 |
Rapid Assessment of PCR Product Quality and Quantity | 39 |
Southern Blotting of Genomic DNA for DNA Profiling | 49 |
Oligonucleotide Probes for SingleLocus and Multilocus VNTR | 57 |
Preparation and Use of 32PLabeled SingleLocus VNTR Probes | 69 |
Statistical Methods Employed in Evaluation of SingleLocus Probe | 83 |
Minisatellite Variant Repeat Unit Mapping Using PCR MVRPCR | 225 |
Sex Determination by PCR Analysis of the XY Amelogenin Gene | 245 |
Recovery of HighMolecularWeight DNA from Blood and Forensic | 1 |
Recovery of DNA for PCR Amplification from Blood and Forensic | 9 |
PCR Analysis of DNA from Fresh and Decomposed Bodies | 19 |
Flaps and Other SalivaStained Material | 27 |
Use of the AmpliType PM + HLA DQA1 PCR Amplification | 261 |
The Use of Capillary Electrophoresis in Genotyping STR Loci | 279 |
SolidPhase Minisequencing as a Tool to Detect DNA Polymorphism | 291 |
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Common terms and phrases
allelic ladder amelogenin AmpliType analysis annealing approx assay automated bands Biotechniques blotting Budowle buffer capillary capillary electrophoresis cells centrifugation Chelex concentration containing cycles denaturation detection DNA extraction DNA fingerprint DNA fragments DNA Profiling Protocols DNA samples DNA sequence dNTP EDTA electrophoresis enzyme ethanol ethidium bromide filter fluorescence Forensic DNA Profiling Forensic Sci gene Genet genomic DNA genotype HLA DQA1 human DNA HUMTH01 hybridization hypervariable Incubate Jeffreys labeled laboratory loading locus membrane Methods in Molecular mg/mL MgCl2 minisatellite minisequencing mitochondrial DNA multilocus multiplex MVR-PCR Note nucleotide oligonucleotide optimization PCR amplification PCR products PCR reaction pipet plate polyacrylamide gels polymerase chain reaction polymorphisms primer procedure proteinase proteinase K reagents remove repeat units room temperature short tandem repeat solution Southern blot stain sterile stored at 20°C STR loci STR system Subheading supernatant template thermal cycler tion tray Tris-HCl tube VNTR wash