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DNA Extraction from macroalgaespecies with high levels of polysaccharide and phenolic components for genetic studies
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Abstract: (4137 Views) |
| Isolation of high quality and molecular weight DNA from macroalgae is usually compromised because ofexcessive contamination of secondary metabolites, disregard of the sampling conditions and their morphological properties. DNA extraction is an essential step for molecular analysis. The main objective of this study is to provide a simple and inexpensive method of isolation of pure DNA from dried blades of macroalgae.DNA extraction was performed usingCTAB, phenol-chloroform, and modified CTAB as well. Also exclude the use of liquid Nitrogen for grinding samples. The changes in our modified procedure are including: ratio of tissue to extraction buffer, optimization of extraction buffer components by increase the concentration of EDTA, b-Mercaptoethanol and adding of PVP and Sarkosyl. Also PCR reaction was performed based on tufA gene.Theratio of A260/A280 and A260/A230 (nm) were recorded between1.7 and 1.84and >2, respectively.Also the concentration of extracted DNA ranged from 6.5 to 8 mgmL-1.Clear band of PCR products were observed in agarose gel.Purity and high quantity of extracted DNA from modified CTAB method was excellent as evident by ratio result in compare with two other methods. Results also suggested that the preparations were sufficiently free of proteins and polyphenolics/polysaccharide compounds which is sufficient for molecular analysis. Polysaccharide and phenolic compound inhibit enzymes of PCR reaction,sosuccess of PCR amplifying confirmed the purity of the product and itsapplicationin molecular analysis. ExtractedDNA from our modified method is suitable in molecular study. |
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| Keywords: DNA extraction, CTAB method, Macroalgae, Polysaccharide components, tufA gene. |
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Full-Text [PDF 385 kb]
(6382 Downloads)
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Type of Study: Research |
Subject:
Fisheries
Received: 2016/04/3 | Accepted: 2016/04/3 | Published: 2016/04/3
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