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

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

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.

• Content Type Journal Article
• DOI 10.1007/s10811-010-9519-5
• Authors
  
  
  • Guangting Wu, State Oceanic Administration Key Laboratory of Marine Bioactive Substances 266061 Qingdao China
  • Chenlin Liu, State Oceanic Administration Key Laboratory of Marine Bioactive Substances 266061 Qingdao China
  • Shenghao Liu, State Oceanic Administration Key Laboratory of Marine Bioactive Substances 266061 Qingdao China
  • Bailin Cong, State Oceanic Administration Key Laboratory of Marine Bioactive Substances 266061 Qingdao China
  • Xiaohang Huang, State Oceanic Administration Key Laboratory of Marine Bioactive Substances 266061 Qingdao China
  
  

• Journal Journal of Applied Phycology
• Online ISSN 1573-5176
• Print ISSN 0921-8971

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