Abstract
To study the molecular mechanism of the Antarctic sea–ice alga in adaptation to polar sea–ice environments, the RNA was prepared for cDNA library construction of Chlamydomonas sp. ICE-L. Three different methods were tested to prepare total RNA from this psychrophilic, unicellular green alga rich in protein and polysaccharide. Lauryl sodium sulfate- based method allowed a most effective extraction of high-quality total RNA compared to the other methods. Total RNA extracted with this protocol was used for cDNA library construction. The recombination rate of constructed cDNA library was 98.60%, the primary titer was 7.15 × 106 pfu, and an average sequence length was 1.2 kb. These results show that with a high-quality RNA preparation, a cDNA library can be constructed successfully for Chlamydomonas sp. ICE-L.
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This work was financially supported by a grant of National Natural Science Foundation (40876106) for Chenlin Liu and grants of polar Stratagem Foundations (2006 and 2007) for Chenlin Liu and Shenghao Liu.
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Wu, G., Liu, C., Liu, S. et al. High-quality RNA preparation for cDNA library construction of the Antarctic sea–ice alga Chlamydomonas sp. ICE-L. J Appl Phycol 22, 779–783 (2010). https://doi.org/10.1007/s10811-010-9519-5
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DOI: https://doi.org/10.1007/s10811-010-9519-5