Isolation of Total RNA from a Freshwater Green Alga, Zygnema cruciatum, Containing High Levels of Pigments

doi:2007.22.2.125

Algae > Volume 22(2); 2007 > Article

Algae 2007;22(2): 125-129. doi: https://doi.org/10.4490/algae.2007.22.2.125

Jong Won Han¹, Minchul Yoon², Key Pyoung Lee¹ and Gwang Hoon Kim²^*

¹Department of Chemistry, Kongju National University, Kongju, 314-701, Korea
²Department of Biology, Kongju National University, Kongju, 314-701, Korea

ABSTRACT

Conventional methods for the isolation and purification of mRNA from Zygnema were unsuccessful because of its high amount of pigments and RNA interactive molecules. In particular, pigments were difficult to remove using conventional protocols because they interacted with RNA during pulverization of the materials. This resulted in total degeneration of RNA in two to three hours. To alleviate this problem, we developed an isolation method that utilized DEAE-cellulose resin. The pigments bound to DEAE anion exchange resin and separated from the RNA. Purified total RNA showed an yield of 50 μg per 100 mg of tissue with this method. The amplified 2nd strand cDNA was distributed 300 bp and over.

Key words: DEAE-cellulose, freshwater green alga, total RNA, Zygnema cruciatum