Rapid and effective method of RNA isolation from green microalga Ankistrodesmus convolutus
The rapid and effective method for the isolation of RNA from green microalga Ankistrodesmus convolutus based on homogenization in a simple CTAB buffer and selective precipitation of RNA with lithium chloride is developed. This procedure avoids the use of toxic chaotropic agents and phenol while...
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| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | en |
| Published: |
Humana Press
2009
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| Online Access: | http://psasir.upm.edu.my/id/eprint/16427/1/Rapid%20and%20effective%20method%20of%20RNA%20isolation%20from%20green%20microalga%20Ankistrodesmus%20convolutus.pdf http://psasir.upm.edu.my/id/eprint/16427/ |
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| Summary: | The rapid and effective method for the isolation
of RNA from green microalga Ankistrodesmus convolutus
based on homogenization in a simple CTAB buffer and
selective precipitation of RNA with lithium chloride is
developed. This procedure avoids the use of toxic chaotropic
agents and phenol while high concentration of
dithiothreitol is used to inhibit RNase activity and prevent
oxidative cross-linking of nucleic acids by phenolics. The
extraction procedure was able to produce high quality and
intact RNA from A. convolutus. The yield of total RNA
was 0.69–0.73 mg/g of fresh weight, with A260/A280 ratio
of 1.79–1.86. The obtained RNA was of sufficient quality
and suitable for downstream application such as RT-PCR
and cDNA library construction. The procedure may also
have wider applicability for total RNA isolation from other
green microalgae species. |
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