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RESEARCH PRODUCT
ABO genotyping by PCR-RFLP and cloning and sequencing
Kurt W. AltJoachim BurgerHaak Wolfgangsubject
Sequence analysisBiologyPolymerase Chain ReactionABO Blood-Group Systemlaw.inventionlawABO blood group systemGenotypeHumansCloning MolecularGenotypingAllelesHistory AncientEcology Evolution Behavior and SystematicsPolymerase chain reactionGeneticsReproducibility of ResultsSequence Analysis DNAGeneral MedicineForensic MedicineRestriction enzymePhenotypeAncient DNAArchaeologyBlood StainsPostmortem ChangesAnthropologyDNA Transposable ElementsAnimal Science and ZoologyChromosome DeletionRestriction fragment length polymorphismToothPolymorphism Restriction Fragment Lengthdescription
A refined PCR-RFLP based method was established to genotype ABO blood groups. The main objective of this study was to make the techniques also suitable for working with degraded DNA. Specific primer design was carried out to choose fragments shorter than 200 bp as necessary in forensic and archaeological applications. Four fragments of exon 6 and 7 of the ABO gene were amplified and digested by in total 7 restriction endonucleases. Particular attention was paid to the base changes at nucleotide positions 261(delG), 297, 526, 703, 721, 771, 796 and 1060(delC) in order to distinguish the six common alleles A101, A201, B, O01, O02 and O03. Furthermore, this method also enables determination of seven of the less frequent alleles: A104, A204, Ax02, Ax03, O05, O06 and O07. The method was applied successfully to a series of blood samples with known phenotypes and genotypes as well as DNA extracted from a thirty year old blood stain and an ancient tooth sample. However, working with ancient DNA requires additional cloning and sequencing of the RFLP-typing results due to DNA post mortem damages such as deaminations, which could lead to false typing results.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2005-01-15 | Anthropologischer Anzeiger |