Automated DNA profiling employing multiplex amplification of short tandem repeat loci.
We have employed automated fluorescence-based technology to detect amplified tri-, tetra-, and pentanucleotide short tandem repeat (STR) loci electrophoresed on denaturing polyacrylamide sequencing gels. The system described incorporates an internal size standard in each sample, allowing the STR-PCR products to be sized automatically with a high degree of precision. By utilizing different fluorescent dye markers for loci that have overlapping allele size ranges, we have developed three multiplex STR systems containing a total of 14 different loci. These multiplex systems were then used to evaluate the usefulness of the 14 loci for the identification of individuals. Allele frequency data were collected from a minimum of 50 individuals from each of three different racial groups: Caucasians, Afro-Caribbeans, and Asians. Of the resulting 42 locus population sets, deviation from Hardy-Weinberg equilibria was detected in only the STR HUMCYARO3-Caucasian data. The probabilities of two unrelated individuals matching by chance (pM) at all 14 loci in the three multiplex reactions was < 1 x 10(14). The combination of multiplex STR-PCR and automatic fluorescence-based detection is thus a rapid and powerful technique for individual identification.
Kimpton
C P
CP
Central Research and Support Establishment, Forensic Science Service, Reading, Berks, United Kingdom.
Gill
P
P
Walton
A
A
Urquhart
A
A
Millican
E S
ES
Adams
M
M
D001322:Autoanalysis / Q000379:methods; D001483:Base Sequence; D002874:Chromosome Mapping; D002877:Chromosomes, Human; D004247:DNA / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005819:Genetic Markers; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D044469:Racial Groups / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid; D013050:Spectrometry, Fluorescence / Q000379:methods
D017931:DNA Primers; D005819:Genetic Markers; D004247:DNA