Investigating Alternative Culture Storage and Obtaining High-Quantity RNA from Chlorella vulgaris

Future space exploration missions will require a nearly closed-loop life support system. Algae are an important model that can be incorporated in these systems for their ability to fixate carbon, recycle water, and as a dietary supplement. One such microorganism, Chlorella vulgaris, has been commonly used in space-based research, but has multiple challenges to overcome before fully understanding the effects of the space environment on this alga can occur. First, maintaining stock cultures in cold storage has typically utilized a nitrogen freezer (-192°C) which can prove impractical as it requires frequent upkeep to manage nitrogen levels and has limited space. Instead, methods utilizing a standard -80°C freezer would become much easier to handle, but research storing C. vulgaris is limited in this condition. The second issue involves low-yield or low-quality RNA extractions from this alga unless a nitrogen freezer is used. This research has two aims: 1) evaluate multiple reagent mixtures to provide an optimal long-term storage option using a -80°C freezer. 2) Extract RNA utilizing various commercially available kits to determine the highest quantity and quality that can be obtained in conjunction with a -80°C freezer