Whole genome amplification strategy for forensic genetic analysis using single or few cell equivalents of genomic DNA.
Evidentiary items sometimes contain an insufficient quantity of DNA for routine forensic genetic analysis. These so-called low copy number DNA samples (< 100 pg of genomic DNA) often fall below the sensitivity limitations of routine DNA analysis methods. Theoretically, one way of making such intractable samples amenable to analysis would be to increase the number of starting genomes available for subsequent STR (short tandem repeat) analysis by a whole genome amplification strategy (WGA). Although numerous studies employing WGA have focused primarily on clinical applications, few in-depth studies have been conducted to evaluate the potential usefulness of these methods in forensic casework. After an initial evaluation of existing methods, a modified WGA strategy was developed that appears to have utility for low copy number forensic casework specimens. The method employs a slight, but important, modification of the "improved primer extension preamplification PCR" method (I-PEP-PCR), which we term mIPEP (modified-I-PEP-PCR). Complete autosomal STR and Y-STR (Y chromosome short tandem repeat) profiles were routinely obtained with 5 pg of template DNA, which is equivalent to 1-2 diploid cells. Remarkably, partial Y- and autosomal STR profiles were obtained from mIPEP-treated DNA recovered from bloodstains exposed to the outside environment for 1 year whereas non-mIPEP-treated samples did not produce profiles. STR profiles were obtained from contact DNA from single dermal ridge fingerprints when the DNA was subjected to prior mIPEP amplification but not when the mIPEP step was omitted.
Hanson
Erin K
EK
Graduate Program in Biomolecular Science, University of Central Florida, PO Box 162366, Orlando, FL 32816-2366, USA.
Ballantyne
Jack
J
D001801:Blood Stains; D001826:Body Fluids / Q000737:chemistry; D004247:DNA / Q000032:analysis*; D005260:Female; D005385:Fingers; D044707:Forensic Sciences / Q000379:methods*; D015894:Genome, Human / Q000235:genetics*; D006801:Humans; D008297:Male; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012661:Semen / Q000737:chemistry; D012680:Sensitivity and Specificity; D012867:Skin / Q000737:chemistry / Q000166:cytology; D014621:Vagina / Q000378:metabolism
D004247:DNA