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  • Whole-Genome Sequencing in Microbial Forensic Analysis of Gamma-Irradiated Microbial Materials.

Whole-Genome Sequencing in Microbial Forensic Analysis of Gamma-Irradiated Microbial Materials.

Applied and environmental microbiology (2015-11-15)
Stacey M Broomall, Mohamed Ait Ichou, Michael D Krepps, Lauren A Johnsky, Mark A Karavis, Kyle S Hubbard, Joseph M Insalaco, Janet L Betters, Brady W Redmond, Bryan A Rivers, Alvin T Liem, Jessica M Hill, Edward T Fochler, Pierce A Roth, C Nicole Rosenzweig, Evan W Skowronski, Henry S Gibbons
摘要

Effective microbial forensic analysis of materials used in a potential biological attack requires robust methods of morphological and genetic characterization of the attack materials in order to enable the attribution of the materials to potential sources and to exclude other potential sources. The genetic homogeneity and potential intersample variability of many of the category A to C bioterrorism agents offer a particular challenge to the generation of attributive signatures, potentially requiring whole-genome or proteomic approaches to be utilized. Currently, irradiation of mail is standard practice at several government facilities judged to be at particularly high risk. Thus, initial forensic signatures would need to be recovered from inactivated (nonviable) material. In the study described in this report, we determined the effects of high-dose gamma irradiation on forensic markers of bacterial biothreat agent surrogate organisms with a particular emphasis on the suitability of genomic DNA (gDNA) recovered from such sources as a template for whole-genome analysis. While irradiation of spores and vegetative cells affected the retention of Gram and spore stains and sheared gDNA into small fragments, we found that irradiated material could be utilized to generate accurate whole-genome sequence data on the Illumina and Roche 454 sequencing platforms.

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Sigma-Aldrich
变溶菌素 来源于球孢链霉菌 ATCC 21553, lyophilized powder, ≥4000 units/mg protein (biuret), Chromatographically purified
Sigma-Aldrich
变溶菌素 来源于球孢链霉菌 ATCC 21553, free of DNA contaminants, suitable for Microbiome research, lyophilized powder, ≥4000 units/mg protein (biuret)