DNA extraction from slug epithelial tissue
Comparison of Different Genomic DNA Extraction Methods in Pest Slugs(Mollusca: Gastropoda)
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The aim of the research was to compare three protocols: Extraction with SDS; Extraction with CTAB and 2¿-mercaptoethanol and Extraction with SDS and proteinase K, to determine the most appropriate method to obtain DNA of high molecular weight and purity in slug epithelial tissue: Arion subfuscus and Deroceras reticulatum. DNA concentration and purity were estimated spectrophotometrically (absorbance at 260 nm and Abs260/Abs280 nm ratio). A completely randomized design with six replicates was used. Significant differences (p
The aim of the research was to compare three protocols: Extraction with SDS; Extraction with CTAB and 2¿-mercaptoethanol and Extraction with SDS and proteinase K, to determine the most appropriate method to obtain DNA of high molecular weight and purity in slug epithelial tissue: Arion subfuscus and Deroceras reticulatum. DNA concentration and purity were estimated spectrophotometrically (absorbance at 260 nm and Abs260/Abs280 nm ratio). A completely randomized design with six replicates was used. Significant differences (p<0.01) were detected for amount, concentration and purity between protocols. The protocol (SDS and proteinase K) allowed obtaining DNA with higher molecular weight (2 514.90±291.56) and purity (1.97±0.02). The quality of the DNA of each protocol was verified by PCR-RAPD, observing little reproducibility for the OPA-02 primer (SDS protocol); while in the other methods high resolution amplification profiles and reproducible bands were observed. It is recommended to use the SDS protocol and proteinase K, to obtain genomic DNA of high molecular weight, quality and purity in slugs.
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