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GENAQUA GENETICS OF AQUATIC ORGANISMS 2017, Vol 1, Num, 1     (Pages: 001-007)

A Quick and Efficient Method for DNA Isolation from Freshwater Ostracods

Nerdin Kubanç 1 ,Tunahan Taşçi 2 ,Cüneyt Kubanç 1 ,Oya Özuluğ 1 ,Vahap Eldem 1

1 Istanbul University, Department of Biology, Faculty of Science, 34134, Istanbul, Turkey
2 Istanbul Bilgi University, Departmant of Medical Imaging Techniques, Vocational School of Health Services, 34387, Istanbul, Turkey
DOI : 10.4194/2459-1831-v1_1_01 Viewed : 394 - Downloaded : 279 Freshwater ostracods are ecologically and evolutionarily important small free-living crustaceans occurring in lakes, rivers and small water bodies. However, despite their importance, only a few genetic studies have been conducted so far in ostracoda due to isolation of in sufficient amounts of DNA. We present a simple, efficient and cost-effective total DNA extraction protocol from freshwater ostracods for PCR amplification. Total DNA was extracted from the four freshwater ostracods; Heterocypris incongruens, Cypridopsis vidua, Eucypris virens and Herpetocypris chevreuxi. We also compared the performance of our protocol with two manual DNA extraction protocols (HotSHOT, Chelex-100) and a commercial kit (Highpure PCR template preparation kit-Roche). The success of the isolation protocol was tested by PCR and sequencing of 18S rRNA and COI regions. Although our technique, Roche and HotSHOT methods resulted in acceptable DNA concentrations and showed an amplifiable band of expected size, both our protocol and Roche generally give better results than HotSHOT, whereas the poor results were obtained from Chelex protocol. Therefore, we suggest that filter-based DNA isolation is more reproducible than other protocols. Our protocol requires no hazardous organic solvents and adaptable to 0.5 mL 8-strip PCR tubes format facilitating DNA extraction from a large number of samples. Keywords : Total DNA, Freshwater ostracods, PCR