Efficient and Cost-Effective Extraction of Genomic DNA From Formalin-Fixed and Paraffin-Embedded Tissues

@article{Weiss2011EfficientAC,
  title={Efficient and Cost-Effective Extraction of Genomic DNA From Formalin-Fixed and Paraffin-Embedded Tissues},
  author={Alexander Th. A. Weiss and N M Delcour and Anja Meyer and Robert Klopfleisch},
  journal={Veterinary Pathology},
  year={2011},
  volume={48},
  pages={834 - 838}
}
Diagnostic and investigative molecular pathology frequently has to resort to extraction of DNA from formalin-fixed and paraffin-embedded tissue samples. Although many different protocols are reported for this type of material, extraction of sufficient amounts of intact DNA is still challenging. Here, the authors report a reproducible, simple, cost-effective, and efficient protocol that yields up to 140 μg of DNA from approximately 10 to 15 mg of formalin-fixed and paraffin-embedded tissue… 

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References

SHOWING 1-9 OF 9 REFERENCES

Efficiency evaluation of a DNA extraction and purification protocol on archival formalin-fixed and paraffin-embedded tissue.

Evaluation of DNA extraction methods and real time PCR optimization on formalin-fixed paraffin-embedded tissues.

This study indicates that DNA obtained from formalin fixed paraffin embedded tissue is highly fragmented and can be used for successful amplification of shorter amplification products up to 250 bp in length.

Comparison of methods in the recovery of nucleic acids from archival formalin-fixed paraffin-embedded autopsy tissues.

The Isolation of Nucleic Acids from Fixed, Paraffin-Embedded Tissues–Which Methods Are Useful When?

This study investigates the quality of nucleic acids recovered from a test panel of fixed specimens that have been manipulated following a number of the published protocols, and provides a quick reference table that can be used to determine appropriate protocols for particular aims.

Effect of fixatives and tissue processing on the content and integrity of nucleic acids.

Characterization of Feline Immunoglobulin Heavy Chain Variable Region Genes for the Molecular Diagnosis of B-cell Neoplasia

Using this assay, FFPE normal feline lymphoid tissues and unfixed peripheral blood mononuclear cells were determined to be composed of polyclonal populations of B cells.