Recovery and STR amplification of DNA from RFLP membranes

S.A. Steadman, J.D. McDonald, J.S. Andrews, N.D. Watson

Research output: Contribution to journalArticle

7 Citations (Scopus)

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.
LanguageEnglish
Pages349-358
Number of pages10
JournalJournal of Forensic Sciences
Volume53
Issue number2
DOIs
Publication statusPublished - Mar 2008

Fingerprint

Restriction Fragment Length Polymorphisms
Membranes
DNA
Polymerase Chain Reaction
Nylons
Detergents
Microsatellite Repeats
Buffers
Coloring Agents
Technology
Acids

Keywords

  • forensic science
  • DNA typing
  • polymerase chain reaction
  • restriction fragment length polymorphism
  • membrane
  • amplification

Cite this

Steadman, S. A., McDonald, J. D., Andrews, J. S., & Watson, N. D. (2008). Recovery and STR amplification of DNA from RFLP membranes. Journal of Forensic Sciences, 53(2), 349-358. https://doi.org/10.1111/j.1556-4029.2008.00674.x
Steadman, S.A. ; McDonald, J.D. ; Andrews, J.S. ; Watson, N.D. / Recovery and STR amplification of DNA from RFLP membranes. In: Journal of Forensic Sciences. 2008 ; Vol. 53, No. 2. pp. 349-358.
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Steadman, SA, McDonald, JD, Andrews, JS & Watson, ND 2008, 'Recovery and STR amplification of DNA from RFLP membranes' Journal of Forensic Sciences, vol. 53, no. 2, pp. 349-358. https://doi.org/10.1111/j.1556-4029.2008.00674.x

Recovery and STR amplification of DNA from RFLP membranes. / Steadman, S.A.; McDonald, J.D.; Andrews, J.S.; Watson, N.D.

In: Journal of Forensic Sciences, Vol. 53, No. 2, 03.2008, p. 349-358.

Research output: Contribution to journalArticle

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