Abstract
In formalin-fixed and paraffin wax-embedded tissues of necropsy origin only short sequences can be amplified (usually less than 100 bases), because of the DNA degradation. This chapter provides a simple treatment to partially restore highly degraded DNA from formalin-fixed paraffin-embedded tissues in order to be able to amplify longer fragments. The method is based on a pre-PCR restoration step to fill the single strand breaks and on a simple denaturation step. No modification to the commonly used extraction procedure is needed. By the use of this simple procedure it is possible to amplify stretches of up to 300 bases even in autopsy FFPE tissues.
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Notes
- 1.
Clean the pipettes with alcohol or another disinfectant and leave them under the UV lamp for almost 10 min. Alternatively, it is possible to autoclave the pipettes according to the provider instructions.
References
Bonin S, Petrera F, Niccolini B, Stanta G (2003) PCR analysis in archival postmortem tissues. Mol Pathol 56(3):184–186
Bonin S, Petrera F, Rosai J, Stanta G (2005) DNA and RNA obtained from Bouin’s fixed tissues. J Clin Pathol 58(3):313–316
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© 2011 Springer-Verlag Berlin Heidelberg
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Bonin, S., Tavano, F. (2011). Restoration and Reconstruction of DNA Length. In: Stanta, G. (eds) Guidelines for Molecular Analysis in Archive Tissues. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-17890-0_11
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DOI: https://doi.org/10.1007/978-3-642-17890-0_11
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Publisher Name: Springer, Berlin, Heidelberg
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