An efficient protocol for genomic DNA extraction from formalin-fixed paraffin-embedded tissues

作者：Santos Sara; Sa Daniela; Bastos Estela; Guedes Pinto Henrique; Gut No; Gartner Fatima; Chaves Raquel^*

来源：Research in Veterinary Science, 2009, 86(3): 421-426.

DOI：10.1016/j.rvsc.2008.08.007

• DNA extraction
• Formalin-fixed paraffin-embedded tissues
• Cat mammary tumours
• PCR amplification
• Sequencing

Formalin-fixed paraffin-embedded tissues (FFPET) represent the largest source of archival biological material available for genomic studies. In this work we present an advanced protocol for extraction of high quality DNA from FFPET that can be applied in several molecular studies. Although cat mammary tumours (CMT) are the third most frequent turnout in cats the recovery of significant number of samples for molecular studies are in some way restricted to FFPET samples. We were able to obtain high quality DNA from FFPET of thirty six CMT that were subjected to pre-fixation and fixation processes routinely used in the veterinary hospitals. The quality of DNA obtained was tested by PCR amplification using six sets of primers that amplify single-copy fragments. The DNA fragments obtained were further sequenced. This protocol was able to provide FFPET gDNA that can be amplified and sequenced for larger fragments up to 1182 bp.

• 出版日期2009-6
• 单位中国地震局