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Recovery and STR Amplification of DNA from RFLP Membranes

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Abstract: 

Restriction fragment length polymorphism (RFLP) techniques were utilized in the forensic DNA community until the mid 1990s when less labor-intensive polymerase chain reaction short tandem repeat (PCR STR) techniques became available. During the transition from RFLP technology to PCR-based STR platforms, a method for comparing RFLP profiles to STR profiles was not developed. While the preferred approach for applying new technology to old cases would be to analyze the original biological stain, this is not always possible. For unsolved cases that previously underwent RFLP analysis, the only DNA remaining may be restriction cut and bound to nylon membranes. These studies investigate 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. 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.
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Keywords: DNA typing; amplification; forensic science; membrane; polymerase chain reaction; restriction fragment length polymorphism

Document Type: Research Article

Affiliations: 1: Sedgwick County Regional Forensic Science Center, 1109 North Minneapolis, Wichita, KS 67214. 2: Wichita State University, 1845 Fairmount, Wichita, KS 67260. 3: University of North Texas Health Science Center, 3500 Camp Bowie Blvd, Fort Worth, TX 76107. 4: University of Strathclyde, 16 Richmond Street, Glasgow G1 1XW, United Kingdom.

Publication date: March 1, 2008

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