A DNA Extraction Method Applied for Living Seahorses
The DNA extraction is the first step provided DNA samples for further genetic studies. As a small animal the DNA extraction from seahorse means killing the living specimens. This study is investigated to estimate the tissue sample size which can be used for extraction of enough DNA for further studies without killing living specimens and find out the suitable method for extraction of DNA from seahorse. Tissue samples were obtained by taking small tail rings at the end of the tails or the fins of living seahorses. Two methods were applied for extraction of DNA: Phenol method and salt method. The extracted DNA was concentrated by Nanodrop system. The quality of DNA was measured by agarose gel and PCR assay. While DNA from fin tissue gave no result in PCR, all DNA from tail tissue with about 20ng/ul gave the good PCR result even extracted by phenol or salt method. With the tail tissue, high concentration of DNA was achieved in both methods. 41% samples and 57.4% samples gave DNA concentration higher than 20ng/ul by phenol and salt method, respectively. The purity of DNA yield extracted by phenol is lower than by salt method (18% in phenol method compared to 44.5% in salt method). Thus, salt method gave higher efficiency than phenol method. The salt method gave higher percentage of sample with high concentration and high purity of DNA than phenol method. The salt method is considered as the suitable method. With the weight of tail tissue around 20mg total amount of DNA at least is 3μg which adequate for further genetic analysis. Beside the over success of salt method compare to phenol method, the salt method is lower cost and less hazardous chemicals as in salt method phenol, an expensive chemical and toxic, is not required.
KeywordsSeahorse DNA extraction Phenol method salt method
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