( ctropherograms and allowing identificat...)Fully automated DNA purification and efficient multiplex PCR for analysis ,,, of microsatellite loci

ctropherograms and allowing identification of individual genotypes (see figures "Highly Reproducible Results from Replicate Samples" and "Accurate Microsatellite Analysis of Genomic DNA from Different Donors"). All 6 targets were well amplified from all DNA samples, irrespective of whether the locus was homozygous (both alleles generate a fragment of the same size) or heterozygous (fragments of different size are generated from each allele).

Satisfactory results were obtained using the same number of PCR cycles and the same amount of PCR product for each sample, indicating that minor differences in DNA concentration do not significantly influence multiplex PCR of microsatellite loci.

Reproducible DNA Yields from Whole Blood

Genomic DNA was isolated from ninety-six 200 l whole blood samples (8 replicates from 12 different donors AL) using the QIAamp DNA Blood BioRobot MDx Kit and the BioRobot MDx workstation. Yields varied depending on the cell content of blood from each donor.

Highly Reproducible Results from Replicate Samples

Multiplex PCR analysis of microsatellite loci. Equal volumes (1 l) of DNA preparations were used as template for a 6plex PCR assay using the QIAGEN Multiplex PCR Kit. Fluorescently labeled PCR products were detected using an ABI PRISM 377 Sequencer. Data for the four 6-FAMlabeled PCR products are shown. Homogenous signals were obtained from 4 different DNA preparations from the same donor.


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