Algae. 2007; 22(2): 125-129.
Isolation of Total RNA from a Freshwater Green Alga, Zygnema cruciatum, Containing High Levels of Pigments
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
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.
Keywords : DEAE-cellulose, freshwater green alga, total RNA, Zygnema cruciatum