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NCJ Number: NCJ 236690   Add to Shopping cart   Find in a Library
Title: Double Strand Break Repair of Highly Damaged DNA
Author(s): Jack Ballantyne Ph.D.
Date Published: 2011
Page Count: 98
Sponsoring Agency: National Institute of Justice
US Department of Justice
Office of Justice Programs
United States of America
Grant Number: 2006-DN-BX-K005
Sale Source: National Institute of Justice/NCJRS
Box 6000
Rockville, MD 20849
United States of America

NCJRS Photocopy Services
Box 6000
Rockville, MD 20849-6000
United States of America
Document: PDF 
Type: Report (Study/Research)
Language: English
Country: United States of America
Annotation: This project attempted to repair “in the test tube” highly fragmented genomic DNA in order to retrieve a DNA profile from otherwise intractable environmentally compromised samples.
Abstract: After attempting various procedures that used current technology, researchers concluded that such technology cannot reconstitute two contiguous fragments of DNA to bring it back to its native state after environmental conditions have compromised the samples. The efforts determined that the sequence complexity of genomic DNA poses an insurmountable computation barrier to reconstituting two contiguous fragments of DNA back to the native state, which is required for the successful repair of double strand breaks. Two general strategies were used in an attempt to repair a double strand break. The “molecular biology” approach involved capturing repair substrates that composed oligonucleotides that subtended a SNP locus of interest and whose function was to capture the appropriate genomic fragments from the degraded sample. The successful repair of oligonucleotide substrates that mimicked some SNP loci was obtained by using capture oligonucleotides; however, all subsequent studies that involved fragmented genomic DNA failed to show any signs of repair. A second strategy used the “biochemical” approach, which recapitulated the cell’s biochemical machinery for double-strand break repair in the test tube, specifically non-homologous end joining (NHEJ). NHEJ also yielded successful repair of oligonucleotide substrates that mimicked some SNP loci; however, all subsequent studies that involved fragmented genomic DNA failed to show any signs of successful NHEJ repair. An alternative but related biochemical pathway, MMEJ (Microhomology-Mediated End-Joining Pathway), was reconstituted, but also failed to repair genomic DNA. Instead of direct repair, researchers also developed and tested a modified oligonucleotide ligation assay (OLA) that was designed to recover profiles from fragmented DNA without actually repairing the DNA. This approach also failed. 23 figures, 5 tables, 30 references, reviewers’ comments, and a listing of presentations in which project results were presented
Main Term(s): Police policies and procedures
Index Term(s): Evidence collection ; Trace evidence ; Evidence identification and analysis ; Victim identification ; Suspect identification ; Forensics/Forensic Sciences ; Investigative techniques ; DNA fingerprinting ; NIJ final report
   
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https://www.ncjrs.gov/App/Publications/abstract.aspx?ID=258705

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