Rapid and Effective Method of RNA Isolation from Green Microalga Ankistrodesmus convolutus

• Published in: Molecular biotechnology 2009-10, Vol.43 (2), p.148-153
• Main Authors: Thanh, Tran, Omar, Hishamuddin, Abdullah, Mohd Puad, Chi, Vu Thi Quynh, Noroozi, Mostafa, Ky, Huynh, Napis, Suhaimi
• Format: Article
• Language: English
• Subjects: Algae; Ankistrodesmus convolutus; Biochemistry; Biological and medical sciences; Biological Techniques; Biotechnology; Cell Biology; Chemical Precipitation; Chemistry; Chemistry and Materials Science; Chlorophyta - chemistry; Chlorophyta - genetics; Fundamental and applied biological sciences. Psychology; Human Genetics; Lithium; Lithium Chloride - chemistry; Microalgae; Nucleic acids; Phenols; Protein Science; Ribonucleic acid; RNA; RNA, Algal - chemistry; RNA, Algal - isolation & purification; RNA, Algal - metabolism; Studies
• ISSN: 1073-6085; 1559-0305
• DOI: 10.1007/s12033-009-9182-8

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.