Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.538802
Title: Investigation into methods for recovery and STR typing of limited quantity and membrane bound forensic samples
Author: Steadman, Shelly
ISNI:       0000 0004 2706 0134
Awarding Body: University of Strathclyde
Current Institution: University of Strathclyde
Date of Award: 2010
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Abstract:
Challenging samples presented to the casework laboratory drive advancements in technology for generating DNA profiles from specimens of limited quantity and quality. This thesis describes the sensitivity limitations of current typing platforms and explores recent advancements in technology aimed at generating DNA profiles from difficult samples; this potentially could include DNA preserved on archived membranes from RFLP analyses conducted on cases in the past. Methods investigated include the utility of the GenomiPhi™ whole genome amplification (WGA) kit, PCRboost enhancement additive, Restorase® DNA polymerase repair kit, and alternate sample processing (cryogenic pulverization) and extraction (paramagnetic) methods. During the course of these studies, the premise of this thesis was shifted to the usefulness of these methodologies in a notorious serial murder case under investigation in the United States known as the BTK (bind, torture, kill) crimes. The ongoing investigation of this prolific serial killer evolved into the systematic development of a procedure to recover DNA from archived RFLP membranes for subsequent STR PCR typing. Initial studies revealed that the WGA, PCRboost, Restorase® DNA polymerase, cryogenic pulverization and paramagnetic extraction would offer limited advantage over already validated processes. Therefore, the investigation progressed to testing numerous extraction buffers for removal of DNA from nylon. Lanes excised from non-irradiated membranes had slightly higher yields than cross-linked membrane lanes, while cross-linked lane yields varied between labs. This indicates storage conditions may play a larger role in recovery potential than initially suspected. Anecdotal accounts suggest the existence of other cases where the generation of CODIS compatible profiles from RFLP membranes may be useful. Therefore, demonstrating success in achieving partial STR profiles from archived RFLP membranes opens another possibility in the investigation and prosecution of cold cases.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.538802  DOI: Not available
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