Isolation of Total RNA from Navicula salinicola NLA for Transkriptomic Applications Pradina Sofianti, Lifana, Alfredo Kono, Zeily Nurachman, Yanti Rachmayanti*
Biochemistry Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung
Jl Ganesha 10, Bandung, 40132, Indonesia
*corresponding author: rachmayanti[at]itb.ac.id
Abstract
Navicula salinicola NLA is a species of unicellular microalgae that belongs to the diatom class (Bacillariophyceae) which has potential for biodiesel due to its lipid content. Knowledge of lipid metabolism pathways can help in efforts to increase lipid production by cells. Cell lipid metabolism can be understood through the information stored in datasets of mRNAs or transcriptomes. The aim of this study was to obtain total RNA isolates from the diatom Navicula salinicola NLA which qualified for transcriptomic analysis. The research steps included identifying diatom samples, cultivating cell samples in modified seawater medium, constructing cell growth curves, harvesting cell biomass from cultures in the exponential and stationary phases, total lipid extraction and total RNA isolation from both logarithmic and stationary phase cell biomass and quality analysis of RNA isolates. The results showed that the sample cells were identical to Navicula salinicola NLA which was characterized by light brown cell color, boat-like cell shape and bilateral symmetry. Initial cell density was 5,5 x 10^5 cells mL-1, increasing to 3,1 x 10^7 cells mL-1 after 7 days of cultivation. The total lipid content of cell biomass was 14% (w/w) in the logarithmic phase and 28.5% (w/w) in the stationary phase. The total RNA was 396.12 microgram isolated from the logarithmic phase and 605.72 microgram from the stationary phase. The ratio of A260/A280 was greater than 2.0 indicating a high level of purity of RNA isolates. The results of agarose gel electrophoresis showed clear and distinctive bands for RNA isolates.
