Rapid methods for extracting high-quality DNA from shell-boring “Conchocelis”
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Abstract
The genome DNA required for molecular studies of Porphyra has generally been extracted from either blades or free-living “Conchocelis”. However, the main disadvantages are complicated procedures, high cost, and DNA contamination. In this study, we developed a strategy for pure DNA extraction from shell-boring “Conchocelis” and a single-step technique to prepare PCR templates. The results show that contamination of free-living “Conchocelis” with microalgae was eliminated after the filaments had bored into shells. The electrophoretic banding patterns and intensities were similar between materials obtained from free-living “Conchocelis” and shell-boring “Conchocelis”, suggesting that total DNA extraction from the latter was efficient. Molecular evidence demonstrated additional bands in PCR products of internal transcribed spacer (ITS) amplifications prepared from cultures of free-living “Conchocelis” contaminated with microalgae. The additional bands appeared in all three Porphyra yezoensis replicates and one out of three Porphyra haitanensis replicates, but were not detectable in preparations from shell-boring “Conchocelis”. The DNA extracted from shell-boring “Conchocelis” reduced the risk of microalgal contamination and the single-step technique reduced the time required for the preparation of DNA for molecular studies of Porphyra. The single-step method should be useful for PCR amplification of small amounts of materials, large sample sizes, or samples contaminated with microalgae. The technique may also be suitable for other genera of Bangiaceae, providing that shell-boring “Conchocelis” stages are present in their life cycles.
©2010 by Walter de Gruyter Berlin New York
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Articles in the same Issue
- Research articles
- Reproduction and life history of the green alga Prasiola linearis Jao (Trebouxiophyceae, Chlorophyta)
- Coexistence of Ulva rigida and Ulva laetevirens (Ulvales, Chlorophyta) in Venice Lagoon and other Italian transitional and marine environments
- New records of subtidal benthic marine algae from Wake Atoll, Central Pacific
- Palisada flagellifera (Ceramiales, Rhodophyta) from the Canary Islands, Spain: a new record for the eastern Atlantic Ocean based on morphological and molecular evidence
- New records of crustose seaweeds associated with subtidal maërl beds and gravel bottoms in Galicia (NW Spain)
- Rapid methods for extracting high-quality DNA from shell-boring “Conchocelis”
- Purification and partial characterization of the lectin from the marine red alga Tichocarpus crinitus (Gmelin) Rupr. (Rhodophyta)
- Mangrove species distribution and abundance in relation to local environmental settings: a case-study at Tumpat, Kelantan Delta, east coast of peninsular Malaysia
- Short communications
- Recent loss of Gibraltar seagrasses
- Leaf reddening in seagrasses
- The effect of boat propeller scarring intensity on genetic variation in a subtropical seagrass species
