OPTIMIZATION OF DNA EXTRACTION FROM BROWN ALGAE (PHAEOPHYCEAE) BASED ON A COMMERCIAL KIT1
| Autor: | Sophie Tuffet, Alodie Snirc, Thomas Silberfeld, Jian-Sheng Sun, Jacques Bonnet, Annie Tillier |
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| Rok vydání: | 2010 |
| Předmět: | |
| Zdroj: | Journal of Phycology. 46:616-621 |
| ISSN: | 1529-8817 0022-3646 |
| DOI: | 10.1111/j.1529-8817.2010.00817.x |
| Popis: | Large-scale DNA molecular studies require reliable and efficient tools for DNA extractions. However, for some plant species and brown algae, isolation of high-quality DNA is difficult. We developed a novel method for isolating high-quality DNA from the polysaccharide-rich and polyphenol-rich brown algae based on a commercial kit and protocol (Qiagen) by optimizing the lysis step and including a chloroform/isoamyl alcohol supplementary purification step. DNAs from 24 brown algal species extracted using the original and the modified Qiagen protocol were compared for yield, quality, and effectiveness in PCR amplification. There was no significant difference in the yields between protocols. However, a statistically significant increase in DNA purity was obtained with the modified protocol, for which the A260/A280 and A260/A230 absorbance ratios averaged 1.66 ± 0.05 and 1.31 ± 0.01, respectively, compared to 1.37 ± 0.04 and 0.52 ± 0.04 with the original protocol. DNAs extracted by the modified procedure were more successfully amplified by PCR (nuclear, mitochondrial, and chloroplastic regions) than DNAs extracted using the original commercial kit and protocol. Importantly, the modified protocol can be applied in a high-throughput (e.g., 96-well plate) format, allowing a higher efficiency for downstream molecular analysis. In addition, improved DNA quality could increase its stability for long-term storage. |
| Databáze: | OpenAIRE |
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