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High-quality RNA preparation for cDNA library construction of the Antarctic sea-ice alga Chlamydomonas sp. ICE-L.

J Appl Phycol.. 2010-12;  22(6):779-783
Guangting Wu, Chenlin Liu, Shenghao Liu, Bailin Cong, Xiaohang Huang. Key Laboratory of Marine Bioactive Substances, State Oceanic Administration, 266061, Qingdao, China
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摘要

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 X 106 pfu, and an average sequence length was 1.2 kb. These results show that with a high-qual... More

关键词

Sea-ice alga; Chlamydomonas sp. ICE-L; RNA extraction; Lauryl sodium sulfate (SDS); cDNA library construction