The use of probe-based real-time PCR to quantify human nuclear DNA in forensic analysis was firstly described by Andreasson et al. (2002). A 78 bp region of the human retinoblastoma susceptibility gene (RB1), a nuclear-encoded single-copy gene located on chromosome 13, was the target in a multiplex PCR quantification assay that was also designed to amplify an mtDNA target (see autosomal DNA and mtDNA quantification by a duplex PCR assay in Table 4.1). The system has been shown to detect down to nuDNA single copies in the dilution series of the standard curve and has been applied to quantify nuDNA from different forensic specimens such as skin debris, saliva stains, hair and bloodstains. This assay has been used recently to quantify nuDNA in the roots and distal sections of plucked and shed head hairs and also from fingerprints and accessories (Andreasson et al., 2006). However, the choice of RB1 as a real-time quantitative PCR target may not be ideal because the RB1 sequence o» so
Table 4.1 Real-time PCR assays used in forensic genetics for specific quantification of human DNA targets
Single-copy autosomal DNA quantification by singleplex PCR
Single-copy autosomal DNA quantification by duplex PCR
Single-copy X and Y
quantification by singleplex PCR
HUMTH01 (62 bp)
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