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Asian Journal of Animal Sciences
Year: 2017  |  Volume: 11  |  Issue: 5  |  Page No.: 214 - 220

Development of an Efficient Method for Producing High Quality Genomic DNA in Crustaceans

Sharmeen Rahman, Daniel Schmidt and Jane Hughes    

Abstract: Background and Objective: High quality genomic DNA is essential for any Genotyping by sequencing technique (e.g., Restriction Site Associated DNA Sequencing). However, producing high quality genomic DNA from crustacean specimens has been difficult due to rapid degradation of tissue samples. In this study, an effective preservation and subsequent DNA extraction procedure was described for producing high quality genomic DNA in a crustacean shrimp, Paratya australiensis. Materials and Methods: Tissue samples were preserved following three preservation techniques: freezing (-80°C), 100% ethanol and RNAlater. Then DNA was extracted from each type of preserved sample using three different methods namely Econo spin column extraction, CTAB and salt extraction. Results: High quality DNA was produced only through Econo spin column extraction method from tissue samples preserved in 100% ethanol and RNAlater. In this case DNA fragments of >10,000 bp were produced without any smear or sign of degradation. The CTAB and salt extraction methods demonstrated very low quality or degraded DNA for all other types of preserved samples which is unusable for Restriction Site Associated DNA Sequencing (RAD-seq) library preparation. Conclusion: High quality genomic DNA was extracted only through Econo spin column extraction process preserving sample in 100% ethanol or RNAlater. The approach described here is easy, simple and does not require impractical cryopreservation methods.

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