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 × $ 10^{6} $ 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..
| Medienart: |
Artikel |
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| Erscheinungsjahr: |
2010 |
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| Erschienen: |
2010 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:22 |
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| Enthalten in: |
Journal of applied phycology - 22(2010), 6 vom: 15. Apr., Seite 779-783 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Wu, Guangting [VerfasserIn] |
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| Links: |
Volltext [lizenzpflichtig] |
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| BKL: | |
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| Themen: |
CDNA library construction |
| Anmerkungen: |
© Springer Science+Business Media B.V. 2010 |
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| doi: |
10.1007/s10811-010-9519-5 |
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| funding: |
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| 520 | |a 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 × $ 10^{6} $ 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. | ||
| 650 | 4 | |a Sea–ice alga | |
| 650 | 4 | |a sp. ICE-L | |
| 650 | 4 | |a RNA extraction | |
| 650 | 4 | |a Lauryl sodium sulfate (SDS) | |
| 650 | 4 | |a cDNA library construction | |
| 700 | 1 | |a Liu, Chenlin |4 aut | |
| 700 | 1 | |a Liu, Shenghao |4 aut | |
| 700 | 1 | |a Cong, Bailin |4 aut | |
| 700 | 1 | |a Huang, Xiaohang |4 aut | |
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