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Biotechnology and Applied Biochemistry (2000) 32, (121–125) (Printed in Great Britain)

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Automated extraction and amplification of DNA from whole blood using a robotic workstation and an integrated thermocycler

Maarten L. Smit*, Belinda A. J. Giesendorf†, Sandra G. Heil*, Jacqueline A. M. Vet*, Frans J. M. Trijbels* and Henk J. Blom*¹

*424 Department of Paediatrics, University Hospital Nijmegen (St Radboud), P.O. Box 9101, 6500 HB, Nijmegen, The Netherlands, and †564 Department of Clinical Chemistry, University Hospital Nijmegen (St Radboud), P.O. Box 9101, 6500 HB, Nijmegen, The Netherlands

Abbreviations used: MTHFR, methylenetetrahydrofolate reductase.

¹ To whom correspondence should be addressed (e-mail H. Blom@ckslkn.azn.nl).

Growing knowledge of the genetic basis of inheritable diseases has resulted in a rapidly increasing demand for DNA mutation analysis. Current methods are reliable and suitable for low-throughput mutation analyses, but are unable to cope with the increasing demand for genetic analyses, necessitating the development of new, fully automated and reliable methods. We developed a semi-automated method for DNA mutation analysis by integrating a thermocycler into a robotic pipetting workstation. DNA was extracted from 84 samples of 10 µl of EDTA-treated whole blood using magnetic beads within 2 h. Directly after isolation, the DNA was automatically transferred to an integrated thermocycler for amplification. Our semi-automated method proved to be reliable and robust, showing unambiguously interpretable PCR signals without occurrence of contamination. It is also faster than conventional manual methods. Only a brief manual intervention is required to remove and refit the seal of the PCR plate. This semi-automated assay is a step forward in the development of fully automated assays for DNA mutation analysis.

Received 26 May 2000/3 July 2000; accepted 10 July 2000

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