title: Automation of large-scale HLA oligotyping using a robotic workstation.
abstract: The ability to amplify specific genomic segments by the PCR has made the analysis of DNA polymorphism with oligonucleotidic probes a practical approach to HLA class II typing. However, to make this method more accurate and applicable to large-scale typing, technical and logistic limitations must be overcome and the risk of human errors must be reduced. To address this problem, we developed an automated procedure, using the Biomek 1000. The system consists of a workstation with a robotic arm and an instrument tablet, an electronic interface unit and an IBM PC. It was modified by the addition of a dot-blot apparatus. Automatic preparation of 96 samples for simultaneous DNA amplification is possible, together with PCR product dilution, distribution and dot-blotting. The robot prevents sample contamination, eliminates human errors and reduces operating costs by decreasing the working time by 50%. Thus, the system is suitable for routine analysis, and it also improves typing accuracy. Furthermore, this system can be adapted for a variety of new DNA-typing strategies that require an initial DNA amplification step.
authors.affiliation: INSERM U 25-Hôpital Necker, Paris, France.
authors.forename: S
authors.initials: S
authors.lastname: Caillat-Zucman
authors.forename: H J
authors.initials: HJ
authors.lastname: Garchon
authors.forename: F
authors.initials: F
authors.lastname: Costantino
authors.forename: S
authors.initials: S
authors.lastname: Cot
authors.forename: J F
authors.initials: JF
authors.lastname: Bach
mesh_terms: D000483:Alleles; D004247:DNA / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D005802:Genes, MHC Class II; D005838:Genotype; D000949:Histocompatibility Antigens Class II / Q000032:analysis / Q000235:genetics*; D006650:Histocompatibility Testing; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D012371:Robotics
chemical_list: D000949:Histocompatibility Antigens Class II; D015345:Oligonucleotide Probes; D004247:DNA