Identification of forensic samples by using an infrared-based automatic DNA sequencer.
We have recently introduced a new protocol for analyzing all core loci of the Federal Bureau of Investigation's (FBI) Combined DNA Index System (CODIS) with an infrared (IR) automatic DNA sequencer (LI-COR 4200). The amplicons were labeled with forward oligonucleotide primers, covalently linked to a new infrared fluorescent molecule (IRDye 800). The alleles were displayed as familiar autoradiogram-like images with real-time detection. This protocol was employed for paternity testing, population studies, and identification of degraded forensic samples. We extensively analyzed some simulated forensic samples and mixed stains (blood, semen, saliva, bones, and fixed archival embedded tissues), comparing the results with donor samples. Sensitivity studies were also performed for the four multiplex systems. Our results show the efficiency, reliability, and accuracy of the IR system for the analysis of forensic samples. We also compared the efficiency of the multiplex protocol with ultraviolet (UV) technology. Paternity tests, undegraded DNA samples, and real forensic samples were analyzed with this approach based on IR technology and with UV-based automatic sequencers in combination with commercially-available kits. The comparability of the results with the widespread UV methods suggests that it is possible to exchange data between laboratories using the same core group of markers but different primer sets and detection methods.
Ricci
Ugo
U
Genetics and Molecular Medicine Unit, A. Meyer Hospital, University of Florence, A. Meyer Hospital, Via Luca Giordano 13, 50132 Florence, Italy. ricciugo@tin.it
Sani
Ilaria
I
Klintschar
Michael
M
Cerri
Nicoletta
N
De Ferrari
Francesco
F
Giovannucci Uzielli
Maria Luisa
ML
D016172:DNA Fingerprinting / Q000379:methods*; D005453:Fluorescence; D005554:Forensic Medicine / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D007211:Indoles; D007259:Infrared Rays; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000295:instrumentation* / Q000379:methods
C427728:IRDye800; D007211:Indoles