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. |
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Contents
retation and Statistical Evaluation of Multilocus | 97 |
Brinkmann 105 | 118 |
sis of Amplified FragmentLength Polymorphisms VNTRSTR | 155 |
fication of Short Tandem Repeat Loci Using | 172 |
al Electrophoretic Methods for Genotyping Amplified STR Loci | 181 |
typing STR Loci Using an Automated DNA Sequencer | 193 |
ation of PCR Fragments by Means of Direct Blotting | 209 |
atellite Variant Repeat Unit Mapping Using PCR MVRPCR | 225 |
letermination by PCR Analysis of the XY Amelogenin Gene | 245 |
es Determination by Analysis of the Cytochrome b Gene | 257 |
f the AmpliType PM + HLA DQA1 PCR Amplification | 267 |
Ise of Capillary Electrophoresis in Genotyping STR Loci | 279 |
Phase Minisequencing as a Tool to Detect DNA Polymorphism | 297 |
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Common terms and phrases
agarose alleles allelic ladder amelogenin AmpliType analysis approx automated bands Biotechniques blotting Budowle buffer capillary capillary electrophoresis cells centrifugation Chelex concentration containing cycles databases denaturation detection DNA extraction DNA fingerprint DNA fragments DNA polymerase DNA Profiling Protocols DNA samples DNA sequence dNTP EDTA electrophoresis enzyme ethanol ethidium ethidium bromide filter fluorescence Forensic DNA Profiling Forensic Sci formamide gene Genet genomic DNA genotype GYPA HLA DQA1 human DNA hybridization hypervariable incubate labeled laboratory lane loading locus membrane Methods in Molecular mg/mL minisatellite mixture multiplex MVR-PCR Note nucleotide oligonucleotide optimal PCR amplification PCR products pellet Perkin-Elmer pipet polymerase chain reaction polymorphic primers procedure proteinase K reagents remove repeat units room temperature short tandem repeat stain sterile stored at 20°C STR loci STR system Subheading tandem repeat template thermal cycler tray Tris-HCl typing VNTR wash