Summary
A simple, practical approach for the extraction of PCR-amplifiable DNA for the HLA-DQa gene from bloodstains deposited on various substrates is described. DNA from bloodstains is purified using Chelex 100 ion-exchange resin and then amplified. If amplification is not achieved, the extract is washed through a Centricon 100 dialysis/concentration tube. If the second amplification of this extract produces a negative result, the extract is processed with Chelex 100 again. This approach has been found to be reliable, safe, efficient and economical.
Zusammenfassung
Eine einfache DNA-Extraktionsmethode zur direkten Amplifikation des HLA-DQa-Locus aus auf verschiedenen Spurenträgern angelegten Blutspuren wird beschrieben. Die Extraktion erfolgt mittels des Chelatbildners Chelex 100. Ist die Amplifikation inhibiert, wird das Extrakt in Centricon 100 Mikrokonzentrator-Röhrchen dialysiert. Bleibt auch eine zweite Amplifikation negativ, erfolgt eine erneute Reinigung mittels Chelex 100. Diese Vorgangsweise erwies sich als effizient, zuverlässig und praktikabel.
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Jung, J.M., Comey, C.T., Baer, D.B. et al. Extraction strategy for obtaining DNA from bloodstains for PCR amplification and typing of the HLA-DQa gene. Int J Leg Med 104, 145–148 (1991). https://doi.org/10.1007/BF01369719
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DOI: https://doi.org/10.1007/BF01369719