Optimization of DNA isolation and amplification protocol for Gracilaria and Sargassum species of Tamil Nadu coast
Chalini, K.; Johnson, M.; Almeida, R.S.; Coutinho, H.D.M. (2021). Optimization of DNA isolation and amplification protocol for Gracilaria and Sargassum species of Tamil Nadu coast. Aquat. Bot. 171: 103377. https://dx.doi.org/10.1016/j.aquabot.2021.103377
In: Aquatic Botany. Elsevier Science: Tokyo; Oxford; New York; London; Amsterdam. ISSN 0304-3770; e-ISSN 1879-1522, more
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| Keywords |
Algae
Gracilaria Greville, 1830 [WoRMS]; Sargassum C.Agardh, 1820 [WoRMS]
Marine/Coastal |
| Author keywords |
Marine algae, Amplification, Molecular, 23s rRNA primer |
| Authors | | Top |
- Chalini, K.
- Johnson, M.
- Almeida, R.S.
- Coutinho, H.D.M.
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| Abstract |
The present study proposes a simple and rapid protocol to isolate DNA from marine algae of the genus Sargassum and Gracilaria. To identify the easy, fast and low cost method, four methods of DNA isolation viz., The Robertson Labs methods, STE Buffer method, PureFast-Plant Genomic DNA purification kit by Helini Biomolecules, Chennai and DNA Isolation by Heat Lysis Method were chosen for the present study. To optimize the DNA isolation methods, the thalli of Sargassum plagiophyllum C. Ag. and Gracilaria Salicornia C.Ag. were employed as a source of DNA. The DNA isolated based on three genomic DNA extraction protocols and by heat lysis method varied with reference to method employed for the isolation. By heat lysis method, the DNA of S. plagiophyllum and G. salicornia was isolated and simultaneously amplified (400 bp) using 23S rRNA gene primers. With these results, the DNA of G. salicornia, G. edulis, G. corticata, G. fergusonii, G. verrucosa, S. aquifolium, S. plagiophyllum, S. polycystum, S. tenerrimum and S. swartzii were isolated and amplified using heat lysis method. The heat lysis method is thereby proven to be efficient, rapid, simple, cost effective and requires only a small fraction of the sample. The heat lysis method can be a successful replacement for the conventional DNA isolation methods, when used specifically for DNA barcoding technique targetting organelle DNA. |
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