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Asian Journal of Animal and Veterinary Advances

Year: 2011 | Volume: 6 | Issue: 9 | Page No.: 958-965
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Authors

Wu-Yi Liu

Ke-Jun Zhang

Keywords

Short Communication

Rapid Salt-Extraction of Genomic DNA from Formalin-Fixed Toad and Frog Tissues for PCR-Based Analyses

Wu-Yi Liu and Ke-Jun Zhang
The technical procedures for extraction of DNA from formalin-fixed tissues include many steps such as chemical treatment, enzymatic digestion, phenol-chloroform purification and alcohol precipitation. Formalin-fixed specimens used in molecular cell and DNA studies have shown shortcomings with respect to the efficacy of DNA isolation and subsequent PCR (Polymerase Chain Reaction) amplification. This study was designed to simplify and maximize recovery of PCR-amplifiable DNA from formalin-fixed toad and frog specimens and also to minimize co-extraction of substances that inhibit PCR amplification. This is achieved by a combination of DNA extraction from formalin-fixed muscle tissues using a salt-out buffer consisting of EDTA and proteinase K and NaCl. All steps are performed at room temperature (20-25°C), thereby reducing further degradation of the already damaged fragile specimen DNA and providing an optimal trade-off between DNA release and degradation. The salt-extraction method of genomic DNA presented here allows DNA isolation from formalin-fixed tissues with a minimum of working steps and equipment and rapidly yields much DNA.
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