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NCJRS Abstract

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NCJ Number: 222346 Find in a Library
Title: Recovery and STR Amplification of DNA From RFLP Membranes
Journal: Journal of Forensic Sciences  Volume:53  Issue:2  Dated:March 2008  Pages:349-358
Author(s): Shelly A. Steadman M.S.; J. David McDonald Ph.D.; John S. Andrews M.S.; Nigel D. Watson Ph.D.
Date Published: March 2008
Page Count: 10
Type: Report (Study/Research)
Format: Article
Language: English
Country: United States of America
Annotation: Because many unsolved cases were carried over from the mid-1990s transition from DNA profiles developed by restriction fragment length polymorphism (RFLP) techniques to DNA profiles developed by the less labor-intensive polymerase chain reaction short tandem repeat (PCR STR) techniques, it is sometimes necessary to compare RFLP profiles to STR profiles; the studies described in this report investigated several methods for obtaining STR profiles from archived membrane-bound DNA, the condition of samples tested with RFLP.
Abstract: A partial STR profile was obtained from DNA that was recovered from an archived membrane using conventional extraction buffer components, indicating promise for recovering useful STR information from RFLP membranes that have been maintained in long-term frozen storage. This raises the possibility of recovering DNA from archived membranes and generating STR profiles that could be compared directly to suspect/victim standards or possibly entered into CODIS for database comparison. Although modified versions of casework stain extraction buffer may be sufficient for recovering amplifiable template, more vigorous methods may be needed, depending on the materials and protocols used for the RFLP analysis and the nature of the storage conditions thereafter. The studies investigated several methods for obtaining STR profiles from membrane-bound DNA, including removal of bound DNA with bases, acids, detergents, various chemicals, and conventional cell extraction solutions. Direct multiplex STR amplification of template in the membrane-bound state was also explored. In addition to detailed descriptions of the various methods used in an effort to obtain STR profiles from membrane-bound DNA, this report describes DNA extraction, quantitation, amplification, and analysis; STR amplification of HaeIII-restricted DNA and amplicon sequence analysis, and binding DNA to nylon membrane. 3 tables, 1 figure, and 35 references
Main Term(s): Criminology
Index Term(s): DNA fingerprinting; Forensic sciences; Investigative techniques
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