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Titolo:
PRODUCING STR LOCUS PATTERNS FROM BLOODSTAINS AND OTHER FORENSIC SAMPLES USING AN INFRARED FLUORESCENT AUTOMATED DNA SEQUENCER
Autore:
ROY R; STEFFENS DL; GARTSIDE B; YANG GY; BRUMBAUGH JA;
Indirizzi:
LI COR INC,DIV BIOTECHNOL,POB 4000 LINCOLN NE 68504 NEBRASKA STATE PATROL CRIMINALIST LAB LINCOLN NE 00000 LI COR INC,DIV BIOTECHNOL LINCOLN NE 68504
Titolo Testata:
Journal of forensic sciences
fascicolo: 3, volume: 41, anno: 1996,
pagine: 418 - 424
SICI:
0022-1198(1996)41:3<418:PSLPFB>2.0.ZU;2-C
Fonte:
ISI
Lingua:
ENG
Soggetto:
TETRANUCLEOTIDE REPEAT POLYMORPHISM; POLYMERASE CHAIN-REACTION; PRIMER; PCR; AMPLIFICATION; PROFILES;
Keywords:
FORENSIC SCIENCE; DNA; SHORT TANDEM REPEATS; POLYMERASE CHAIN REACTION; TTH POLYMERASE; INFRARED; AUTOMATED DNA SEQUENCER; FLUORESCENCE; ACTBP2; D2S436; HUMTHO1; D20S470; D18S535;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
23
Recensione:
Indirizzi per estratti:
Citazione:
R. Roy et al., "PRODUCING STR LOCUS PATTERNS FROM BLOODSTAINS AND OTHER FORENSIC SAMPLES USING AN INFRARED FLUORESCENT AUTOMATED DNA SEQUENCER", Journal of forensic sciences, 41(3), 1996, pp. 418-424

Abstract

Short tandem repeat (STR) analysis is increasingly being used in forensic case analysis because of the large number of STR loci in the human genome and their highly polymorphic nature. An automated DNA sequencer using high sensitivity infrared (IR) fluorescence technology was used to detect STR allele patterns from simulated forensic samples. The amplification strategy used a 19 base pair extension on the 5' end of one of the PCR primers. This sequence is identical to the sequence of a universal M13 Forward sequencing primer which is included in the amplification reaction. Allelic bands were detected by incorporation of the M13 primer-fluorescent dye conjugate into PCR products thus eliminating the need for direct conjugation of fluorescent dye to individual STR primers. By using an IR-based automated DNA sequencer and Tth DNA polymerase, polymorphic STR alleles were detected on-line rapidly and efficiently from bloodstains using only a high temperature incubation to extract DNA from blood cells. Five STR loci were also amplified using Chelex extracted DNA from simulated forensic samples. Multiplexing of three primer pairs in a single PCR mixture for amplification was accomplished using Taq polymerase. This system combines IR fluorescence chemistry and laser technology thus eliminating the need for radioactivity and the gel handling required with silver staining and fluor detection systems. Real-time detection permits immediate visualization of the data and STR alleles are displayed as familiar autoradiogramlike images that can be analyzed by computer. By loading a 64 lane gel twiceand multiplexing with three primer pairs, forensic scientists can type at least three loci from 120 samples in one day.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 30/11/20 alle ore 09:54:51