000 02919nam a2200265 4500
001 390977
090 _a390977
100 _a20120531d2011 y0pory50 ba
101 _aeng
200 _aForensic stain identification by RT-PCR analysis (updated)
_bRecurso eletrónico
_fTrisha Conti, Eric Buel
210 _a[Vermont]
_ced. do a.
_d2011
215 _a1 CD-ROM
_d12 cm
300 _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.
330 _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.
606 _aANÁLISE DE VESTÍGIOS
_9320649
606 _aFLUIDO CORPORAL
_9343188
606 _aADN
_9243660
606 _aBIOLOGIA FORENSE
_91092
700 _aCONTI
_bTrisha
_9356777
701 _aBUEL
_bEric Buel
_9356778
955 _xOferta