02683nam a2200193   4500001000700000100004100007101000800048200011000056210003100166215002000197300029000217330186600507606002902373606002002402606000802422606002102430700001802451701002002469390977  a20120531d2011       y0pory50      ba  aeng  aForensic stain identification by RT-PCR analysis (updated)bRecurso eletrónicofTrisha Conti, Eric Buel  a[Vermont]ced. do a.d2011  a1 CD-ROMd12 cm  aResearch report [prepared by Vermont Forensic Laboratory] and funded by the U.S. Department of Justice - Award nr. 2007-DN-BX-K149. Ficheiro de 408 KB em formato PDF (88 p.). Disponível também em: https://www.ncjrs.gov/pdffiles1/nij/grants/236537.pdf. Acedido a 31 de maio de 2012.  aWith the advent of innovative molecular biological techniques becoming the norm in the forensic laboratory, it is plausible to imagine the eventual replacement of traditional serological testing methods used to identify questioned stains with molecular biological techniques. New tests that are tissue-specific and designed to be multiplexed would yield rapid results on a minimal amount of sample. Such testing could employ mRNA as the tissue-specific determinant by testing for the appropriate tissue-specific mRNAs. Analyses can also be performed to demonstrate that mRNA is relatively stable and can thus be of great use in a wide variety of forensic cases. The nature of this research was to identify mRNA transcripts that will definitively identify the tissue of origin, determine if such transcripts survive the typical environmental insults that forensic samples may encounter, and to develop rapid multiplex assays to assess these molecules using small amounts of sample. A crucial prerequisite to these analyses is the development of a DNA/RNA co-extraction method to minimize sample requirements and eliminate the need for two separate extractions. Through collaboration with Promega, a RNA/DNA co-isolation technique was developed which effectively extracts both nucleic acids of sufficient quality and quantity for downstream real-time PCR and STR analyses. The stability of RNA over time was established using real-time PCR assays. Two separate technologies were used to multiplex assays once candidates were shown to be tissue-specific. The Plexor® One-Step qRT-PCR System was used to design a semen-saliva multiplex assay in collaboration with Promega. Unfortunately, Promega has decided to discontinue development of this assay. Lastly, homebrew TaqMan® assays were developed for semen-sperm identification as well as a brain screening assay.  aANÁLISE DE VESTÍGIOS  aFLUIDO CORPORAL  aADN  aBIOLOGIA FORENSE  aCONTIbTrisha  aBUELbEric Buel