The Benefits of the Absolute Quantifying Method with TaqMan Probeversus Other Methods
Journal Title: UNKNOWN - Year 2015, Vol 4, Issue 2
Abstract
Aim. To evaluate in an experimental way the best extraction method which maximally extracts the sample and avoids the issues or DNA degrading.Material and method.The determination of the blood samples of the external environment is done by using the test Kastle Meyer. The determination of the extracted DNA is done by the method Absolute Quantifying with ABI Prism Real Time-PCR 7000. The complex of the reagents used for this aim is the type Quantifiler® Human DNA Quantification Kit. The software used for the analyses was: ABI PRISM® 7000; Sequence Detection System Software Kit version 1.0 The multiplying of the genetic markers is done by using the PCR method and the use of Thermal Cycler Gene Amp 9700. The complex of the genes used for this method is AmpF£STR® Identifier® PCR Amplification Kit. The analyzed locus are: D7S820, CSF1P0, D18S51, Amelogenin and FGA. The separation and the analyzing of these fragments is done by using the genetic analyzer ABI 310 (Applied Biosystem, P/N: 310-00-200/240-W) and softwer GeneMapper® ID Software v3.2 (Applied Biosystem, P/N: 4338951). Results. In the table we notice that samples are positive for hTERT gene. The negative result of this we think comes from the paper substract used for the collection of the sample, which is not appropriate for the deposit of the epithelial cells of the skin when is touched. Conclusion.The samples have been in different levels of damage of DNA molecules, this marker has shown high stability to environmental factors. the use of Quantifiler® Human DNA Quantification Kit increases the accuracy and eficasity in the discovery of human DNA presence and the determination of its amount wich makes this gene have a primary role compared with other markers used for the same purposes.
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