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Unità operativa di Anatomia Patologica, Azienda Ospedaliera “V. Cervello”, Palermo,Italy

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Unit operativa di Anatomia Patologica, Azienda Ospedaliera V. Cervello , Palermo,Italy - PowerPoint PPT Presentation


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EVALUATION OF AN ALTERNATIVE METHOD FOR DNA EXTRACTION FROM PARAFFIN-EMBEDDED TISSUES FOR PCR AMPLIFICATION. G. Costanzo,T. Mannone, E. Gallo, F. Guddo, F. Raiata, V. Randazzo and A.G. Rizzo. Unità operativa di Anatomia Patologica, Azienda Ospedaliera “V. Cervello”, Palermo,Italy.

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EVALUATION OF AN ALTERNATIVE METHOD FOR DNA EXTRACTION FROM PARAFFIN-EMBEDDED TISSUES FOR PCR AMPLIFICATION.

G. Costanzo,T. Mannone, E. Gallo, F. Guddo, F. Raiata, V. Randazzo and A.G. Rizzo.

Unità operativa di Anatomia Patologica, Azienda Ospedaliera “V. Cervello”, Palermo,Italy

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Formalin-fixed, paraffin-embedded tissues are one of the popular sources of diagnostic materials, the easiest to store and trasport.

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The polymerase chain reaction (PCR) is a method for the in vitro amplification of specific nucleic acid sequences.

The major attraction of the PCR, for research and diagnostic purposes in pathology, is its ability to work on low doses of DNA samples obtained by simple methods of extraction.

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There are more studies comparing different techniques of DNA extraction from paraffin-embedded tissues.

In the present study, we compared two methods:

1) Overnight proteinase K digestion followed by 5% Chelex-100 (BioRad) purification

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Both methods were applied to extract DNA from thirty blocks of various organs:

  • lung
  • breast
  • thyroid
  • spleen
  • colon
  • testicle
  • appendix
  • uterus

Chelex-100 method

BioDiagene method

Successful extraction was determined by the ability to amplify a 268-bp fragment of the house-keeping gene beta-globin.

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CONCLUSION

  • Although we were able to obtain amplificable
  • DNA as Chelex-100 method as BioDiagene
  • method
  • We found two important advantages:
  • BioDiagene is faster than Chelex-100 method, infact, in only 30 minutes the DNA is extracted and ready to use for PCR amplification.
  • BioDiagene requires less material, infact it is sufficient only one 10mm thickness section against 2-10 sections for Chelex-100.
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