Abstract
A method called Sequential Multiplex Amplification (SMA) has been developed whereby a limited amount of DNA extracted from a sample can be reutilized for several single polymerase chain reaction (PCR) amplifications. The method involves recovery of genomic template DNA by microfiltration of PCR-amplified samples. Up to 5 different loci have been typed, each in a single system PCR-based assay, beginning with a test quantity of 5 ng template DNA. Genotypes of the DNA donors were compared with those obtained from individual amplifications and shown to be identical. This could be a useful technique for typing a number of loci from a limited amount of DNa and to recover template DNA from samples previously subjected to PCR. Obviously, when small quantities of template DNA are available, this technique can prove quite useful.
Zusammenfassung
Die Methode der sequentiellen multiplex Amplifikation (SMA) ermöglicht die Wiederverwendung einer limitierten Menge genomischer DNA für mehrere aufeinanderfolgende PCR Amplifikationen. Die Isolierung genomischer DNA aus den PCR-amplifizierten Proben erfolgte durch Mikrofiltration. 5 ng genomischer DNA wurden als Ausgangsmenge eingesetzt und bis zu 5 verschiedene Loci in getrennten PCR-Ansätzen nacheinander untersucht. Die Amplifikationsmuster der sequentiellen multiplex Amplifikation wurden mit denen aus individuellen Amplifikationen verglichen und waren identisch. Diese Technik ermöglicht die Untersuchung verschiedener Loci ausgehend von einer begrenzten Menge an genomischer DNA und kann daher besonders Anwendung finden, wenn geringe Mengen an genomischer DNA für die PCR Amplifikation zur Verfügung stehen.
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Lorente, M., Lorente, J.A., Wilson, M.R. et al. Sequential multiplex amplification (SMA) of genetic loci: A method for recovering template DNA for subsequent analyses of additional loci. Int J Leg Med 107, 156–158 (1994). https://doi.org/10.1007/BF01225604
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DOI: https://doi.org/10.1007/BF01225604