STR typing in forensic genetics has been performed traditionally using capillary electrophoresis (CE); however, CE-based method has some limitations: a small number of STR loci can be used; stutter products, dye artifacts and low level alleles. Massively parallel sequencing (MPS) has been considered a viable technology in recent years allowing high-throughput coverage at a relatively affordable price. Some of the CE-based limitations may be overcome with the application of MPS. The results of the current study showed that the MinElute PCR Purification Kit (Qiagen) was a better size selection method compared with recommended diluted bead mixtures. The library input sensitivity study determined that a wide range of amplicon product (6-200 ng) could be used for library preparation without apparent differences in the STR profile. PCR sensitivity study indicated that 62 pg may be minimum input amount for generating complete profiles. Reliability study results on 24 different individuals showed that high depth of coverage (DoC) and balanced heterozygote allele coverage ratios (ACRs) could be obtained with 250 pg of input DNA, and 62 pg could generate complete or nearly complete profiles. These studies indicate that this STR multiplex system and the Illumina MiSeq can generate reliable STR profiles at a sensitivity level that competes with current widely used CE-based method. (Publisher abstract modified)
Downloads
Similar Publications
- Study of CTS DNA Proficiency Tests with Regard to DNA Mixture Interpretation: A NIST Scientific Foundation Review
- Symptoms Consistent With Shift Work Disorder Are Common Across Groups of First Responders
- Organizational Dis trust Comparing Disengagement Among Former Left-Wing and Right-Wing Violent Extremists