Abstract...
DNA profiles can be obtained from fabrics where a person has made direct contact with clothing. A standard approach is to cut out a section of the fabric and then use a commercially available method to extract and isolate the DNA. Alternative methods to isolate DNA include the use of adhesive tape to remove traces of cellular material from the fabric prior to extraction. We report on a process to obtain full DNA profiles using direct amplification from a range of fabrics. The absence of an extraction step both reduces the opportunity for contamination and reduces the loss of DNA during the extraction process, increasing the sensitivity of the process of generating a DNA profile. The process does not require the use of commercially available extraction kits thus reducing the cost of generating a DNA profile from trace amounts of starting material. The results are in part dependent upon the nature of the fabric used to which the DNA has been transferred.
Origine:fsigenetics.com
Visualizzazione post con etichetta Degraded DNA. Mostra tutti i post
Visualizzazione post con etichetta Degraded DNA. Mostra tutti i post
martedì 10 novembre 2009
Casework testing of the multiplex kits AmpFℓSTR® SEfiler Plus™ PCR amplification kit (AB), PowerPlex® S5 System (Promega) and AmpFℓSTR® MiniFiler™ PCR
Abstract
The STR kits SEfiler Plus™ (D3S1358, FGA, D8S1179, D18S51, D21S11, TH01, VWA, SE33, D2S1338, D16S539, D19S433, and Amelogenin), PowerPlex® S5 System (D18S51, D8S1179, TH01, FGA, Amelogenin) and MiniFiler™ (D13S317, D7S820, Amelogenin, D2S1338, D21S11, D16S539, D18S51, CSF1PO, and FGA) were comparatively tested for their robustness and sensitivity. About fifty stains with highly degraded DNA and little DNA quantity served as examination material (e.g. hair with a telogen root, bones, degraded saliva stains on drinking-vessels and skin cell mixtures).
The PowerPlex® S5 with five German-DNA-database (DAD)-systems and the MiniFiler kit with four topical DAD-systems and further STR markers show reduced amplicon lengths. The SEfiler Plus™ kit represents no MiniSTR multiplex, but contains the nine current DAD-systems and further three systems D2S1338, D16S539, D19S433, which are the potential expansion markers for the German-DNA-database.
We have detected on the basis of our comparative stain investigations, that the SEfiler Plus kit was less sensitive than the PowerPlex® S5 and the MiniFiler kit. The MiniFiler™ and the PowerPlex® S5 kit showed a comparative high sensitivity.
Especially in analysing skin cell mixtures, the MiniFiler kit contained larger differences with regard to the performance of the fluorescent dyes/primer concentration co-ordination than the PowerPlex® S5. The SEfiler Plus kit generated – just as both MiniSTR kits – relative robust typing results, but there appeared an increased sensitivity for “allelic drop-outs” and “imbalances”. Since the SEfiler Plus kit was not planned as MiniSTR concept, “allelic drop-outs” were observed, as expected, more frequent in typing stains with degraded DNA and little DNA quantity, especially in the long PCR products (e.g. D18S51).
Origine: fsigenetics.com
The STR kits SEfiler Plus™ (D3S1358, FGA, D8S1179, D18S51, D21S11, TH01, VWA, SE33, D2S1338, D16S539, D19S433, and Amelogenin), PowerPlex® S5 System (D18S51, D8S1179, TH01, FGA, Amelogenin) and MiniFiler™ (D13S317, D7S820, Amelogenin, D2S1338, D21S11, D16S539, D18S51, CSF1PO, and FGA) were comparatively tested for their robustness and sensitivity. About fifty stains with highly degraded DNA and little DNA quantity served as examination material (e.g. hair with a telogen root, bones, degraded saliva stains on drinking-vessels and skin cell mixtures).
The PowerPlex® S5 with five German-DNA-database (DAD)-systems and the MiniFiler kit with four topical DAD-systems and further STR markers show reduced amplicon lengths. The SEfiler Plus™ kit represents no MiniSTR multiplex, but contains the nine current DAD-systems and further three systems D2S1338, D16S539, D19S433, which are the potential expansion markers for the German-DNA-database.
We have detected on the basis of our comparative stain investigations, that the SEfiler Plus kit was less sensitive than the PowerPlex® S5 and the MiniFiler kit. The MiniFiler™ and the PowerPlex® S5 kit showed a comparative high sensitivity.
Especially in analysing skin cell mixtures, the MiniFiler kit contained larger differences with regard to the performance of the fluorescent dyes/primer concentration co-ordination than the PowerPlex® S5. The SEfiler Plus kit generated – just as both MiniSTR kits – relative robust typing results, but there appeared an increased sensitivity for “allelic drop-outs” and “imbalances”. Since the SEfiler Plus kit was not planned as MiniSTR concept, “allelic drop-outs” were observed, as expected, more frequent in typing stains with degraded DNA and little DNA quantity, especially in the long PCR products (e.g. D18S51).
Origine: fsigenetics.com
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