A new multiplex-PCR comprising autosomal and y-specific STRs and mitochondrial DNA to analyze highly degraded material

Wurmb‐Schwark, Nicole von; Preusse-Prange, A.; Heinrich, A.; Simeoni, Eva; Bosch, Thomas C. G.; Schwark, Thorsten

doi:10.1016/j.fsigen.2008.11.007

Cited by 27 publications

(10 citation statements)

References 38 publications

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“…Primers and PCR conditions were used as described previously. 18 Amplification products were separated and detected on an ABIPrism3130 Genetic Analyzer with ROX500 standard fragments (all Applied Biosystems, Weiterstadt, Germany) in comparison with a self-made allelic ladder. Electrophoresis results were analyzed using the GeneMapper ID Software v3.2 with the respective self-designed bin set.…”

Section: Th01 Typingmentioning

confidence: 99%

Association of TH01 with human longevity revisited

et al. 2011

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The gene tyrosine hydroxylase 1 (TH01) has been suggested as a candidate for human longevity. A previous study has shown an association between longevity and specific alleles of the TH01 short tandem repeat (STR) polymorphism in an Italian population. This STR locus is also widely used in forensic genetics. If the TH01-longevity association could be confirmed in independent samples, this finding would have important ramifications for the use of this polymorphism in a forensic context. In the present study, we sought to replicate the previous association result by investigating 471 long-lived individuals (96-110 years) and 462 younger controls (19-75 years) from Germany. In the analyzed samples, the association between TH01 and longevity was not replicated. However, the obtained TH01 allele frequencies were consistent with published data. We observed considerable differences in the allele distribution between Germans and Italians, in particular with regard to allele 9.3, which displayed a previously undetected decreasing West-East and North-South cline across Europe. The discrepant TH01-longevity association results in Germans and Italians could therefore be due to population-specific effects. This finding highlights the need to take into consideration population genetic data when dealing with association studies.

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Section: Th01 Typingmentioning

confidence: 99%

Association of TH01 with human longevity revisited

et al. 2011

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“…More recently, Wurmb-Schwark et al [21] described the development of a new multiplex reaction to assess the degree of DNA degradation in bones. It involved the amplification of five nuclear DNA loci (amelogenin, TH01, vWA, DYS390 and DYS391), and two mitochondrial DNA hypervariable regions along with a 271-bp fragment from Hydra vulgaris as an IAC to detect PCR inhibition.…”

Section: Introductionmentioning

confidence: 99%

Development of internal amplification controls for DNA profiling with the AmpFℓSTR^® SGM Plus^® kit

et al. 2011

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DNA extracted from forensic samples can be degraded and also contain co-extracted contaminants that inhibit PCR. The effects of DNA degradation and PCR inhibition are often indistinguishable when examining a DNA profile. Two internal amplification controls (IACs) were developed to improve quality control of PCR using the AmpFℓSTR® SGM Plus® kit. The co-amplification of these controls with DNA samples was used to monitor amplification efficiency and detect PCR inhibitors. IAC fragments of 90 and 410 bp (IAC₉₀ and IAC₄₁₀) were generated from the plasmid pBR322 using tailed primers and then amplified with ROX-labelled primers. Co-amplification of IAC₉₀ and IAC₄₁₀ was performed with varying amounts of template DNA, degraded DNA and DNA contaminated with humic acid, heme and indigo dye. Both IAC₉₀ and IAC₄₁₀ were successfully amplified with human DNA without significantly affecting the quality of the DNA profile, even with DNA amounts lower than 0.5 ng. In the presence of inhibitors, the IAC₉₀ signal was still present after all human DNA loci fail to amplify; in contrast, the IAC₄₁₀ signal was reduced or absent at low levels of inhibition. Amplification of the two IACs provided an internal PCR control and allowed partial profiles caused by inhibition to be distinguished from degraded DNA profiles.

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“…IPCs currently used in forensic kits have been designed such that the IPC sequence was checked against GenBank to ensure uniqueness-but the methods of sequence design are not always reported. 36 Non-human sequences have been used for this purpose, such as a portion of Sea Pansy (Renilla reniformis) luciferase gene 37 , a hydra-specific (Hydra vulgaris) actin gene segment 22,23 , or rat DNA. 38 In some commercial kits (e.g.…”

Section: Discussionmentioning

confidence: 99%

“…Our results show that nullomer sequences can be used as an IAC, and as molecular tags and barcodes, successfully integrated into the multiplex PCR reactions of commercially available forensic profiling kits, and used along with PCR for sequencing. The use of IACs (sometimes designated internal positive controls, IPCs) is already commonplace for food and clinical microbiological testing, 19 forensic quantification kits, 20 , forensic human identification kits 21,22,23 , and are particularly useful for detecting PCR inhibitors. 24 For PCR-based tests of food-borne pathogens, the European Standardization Committee has developed guidelines that require the presence of an IAC.…”

Section: Introductionmentioning

confidence: 99%

Safeguarding forensic DNA reference samples with nullomer barcodes

Goswami

Davis

Andersen

et al. 2013

Journal of Forensic and Legal Medicine

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A new multiplex-PCR comprising autosomal and y-specific STRs and mitochondrial DNA to analyze highly degraded material

Cited by 27 publications

References 38 publications

Association of TH01 with human longevity revisited

Association of TH01 with human longevity revisited

Development of internal amplification controls for DNA profiling with the AmpFℓSTR^® SGM Plus^® kit

Safeguarding forensic DNA reference samples with nullomer barcodes

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A new multiplex-PCR comprising autosomal and y-specific STRs and mitochondrial DNA to analyze highly degraded material

Cited by 27 publications

References 38 publications

Association of TH01 with human longevity revisited

Association of TH01 with human longevity revisited

Development of internal amplification controls for DNA profiling with the AmpFℓSTR® SGM Plus® kit

Safeguarding forensic DNA reference samples with nullomer barcodes

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Product

Resources

About

Development of internal amplification controls for DNA profiling with the AmpFℓSTR^® SGM Plus^® kit