PLEX-ID mtDNA Assay
The PLEX-ID mtDNA assay is a quantitative, non-diagnostic assay used to interrogate hypervariable (HV) regions I and II, 15924-16428 and 31-576 respectively. The assay combines PCR with high precision electrospray ionization mass spectrometry to establish and compare the base composition profile for mitochondrial DNA in shed hairs, buccal swabs, cut hairs, aged or degraded tissue, bones or teeth and very small quantities of blood, or other bodily fluids. This assay provides more discriminatory power than traditional Sanger sequencing and also allows for mtDNA mixture interpretation, Figure 1.
| Name | List No. | Quantity | Additional Information |
| PLEX-ID mtDNA v2.0 (PLEX-ID) | 03N38-62 | 10 plates (120 samples) |  |
| PLEX-ID mtDNA v2.0 (T5000) | 03N38-02 | 10 plates (120 samples) |
The PLEX-ID mtDNA v2.0 assay includes:
- Ten 96-well plates with all components, with the exception of DNA template, for PCR and analysis
- Package insert
Positive and negative controls are not included.
There is no specific sample extraction protocol available for this assay; however, fluid handling scripts may be available for the PLEX-ID FH, based upon the assay plate configuration, volume, reagents and sample format required.
The PLEX-ID mtDNA assay v2.0 uses 24 primer pairs multiplexed into eight wells to interrogate HVI and II, 15924-16428 and 31-576 respectively. The assay generates base compositions (Aw, Gx, Cy, Tz). The software allows for base compositions to be reviewed, compared to each other, and converts sequence data into a base composition for comparison.
Figure 1: Effect of length heteroplasmy upon mitochondrial DNA sequencing and the PLEX-ID mitochondrial tiling assay.*
* Base Composition Profiling of Human Mitochondrial DNA Using Polymerase Chain Reaction and Direct Automated Electrospray Ionization Mass Spectrometry, Ionization Mass Spectrometry, Analytical Chemistry, 2009, 81, 7515-7526
Additional Information
- Base Composition Profiling of Human Mitochondrial DNA Using Polymerase Chain Reaction and Direct Automated Electrospray Ionization Mass Spectrometry, T. Hall et. al., Anal. Chem. 2009, 81, 7515–7526
- Validation of mass spectrometry analysis of mitochondrial DNA. B Budowle, et. Al. Forensic Science International: Genetics Supplement Series. 2009. 2:527-528.
- Analysis of DNA forensic markers using high throughput mass spectrometry. S Hofstadler, et. al. Forensic Science International: Genetics Supplement Series. 2009. 2:524-526.
- Comparative analysis of human mitochondrial DNA from World War I bone samples by DNA sequencing and ESI-TOF mass spectrometry. R Howard, et. al. Forensic Science International. 2011. FSIGEN-755.
