Scope and Method of Study. An end-point PCR-based multiplex assay was developmentally validated for use in forensic DNA casework that could simultaneously gender-type and quantify total human and male-only DNA. The developmental validation included the studies that are required by the Revised Validation Guidelines set forth by the Scientific Working Group on DNA Analysis Methods. DNA was extracted by an organic extraction method, or the Hair Extraction Kit. The samples were purified and de-salted using DNA IQ(TM) System with Differex(TM) or CentriconsRTM. The NIST SRM 2372 quantitation standards were purchased from the National Institute of Standards and Technology. Samples were amplified using an ABI 9700 thermal cycler and analyzed using an ABI Prism 310 Genetic Analyzer. The peak height and sizing of alleles was determined using the GeneScanRTM Analysis version 3.1.2 software and the alleles were labeled using a Genotyper RTM version 2.5.2 software macro developed for this research.... down to the tens of picograms ranging up to 50ng/I¼L or more (QuantifilerAr and QuantifilerAr Y Usera#39;s Manual, 2006). ... In light of the average HID test targeting 100pg/I¼L, a dynamic range of this magnitude is not needed for casework processing. ... the 500pg that is recommended to generate a complete HID profile from a commercially available STR typing kit. ... The upper and lower bounds for the y-axis (total input DNA) were put in to show the actual input DNA as estimated by aanbsp;...
|Title||:||Developmental Validation of an Improved Multiplex Assay for Use in Forensic Casework: The Simultaneous Gender-typing and Quantification of Total Human and Male-only DNA Within Forensic Samples|
|Author||:||Jonathan Price Wilson|
|Publisher||:||ProQuest - 2008|