RNAlater Solution can be used to stabilize and protect cellular RNA in intact unfrozen samples through rapid penetration of fresh tissue and deactivation of nucleases. Dr. Gorokhova (Stockholm University, Sweden) designed an experiment to determine the amount of time a microcrustacean specimen can be stored in RNAlater Solution as an alternative to deep freezing of the sample. The study demonstrated that RNAlater Solution successfully preserved both RNA and DNA for up to one month at room temperature, and for up to four months at 5°C.
Preserving Aquatic Samples in the Field
Ecologists are interested in understanding environmental factors that support plankton growth in aquatic ecosystems. This requires the processing and analysis of a large number of samples with low RNA and DNA content for estimating growth and nutritional conditions for these small organisms. Successful preservation and processing of collected samples is therefore critical. However, deep freezing in liquid nitrogen is not convenient for aquatic samples collected on oceanographic cruises and remote field studies. Dr. Gorokhava tested whether
RNAlater Tissue Collection: RNA Stabilization Solution could be used for sample preservation, instead of deep freezing for preserving nucleic acids in brine shrimp nauplii* (Artemia salina). Newly hatched shrimp nauplii were stored at (1) -80°C, (2) in RNA
later Solution at 5°C, or (3) in RNA
later Solution at room temperature.
Total cellular RNA and DNA were quantified in individual nauplii using a fluorometric assay. RNA and DNA yields were subsequently compared with control samples (fresh animals assayed immediately after hatching) and among preservation treatments to determine the longterm effects of different sample storage techniques. Nucleic acid quantities were determined for each preservation treatment after one, two, four, and eight months. For both RNA and DNA yields, shrimp samples stored in RNA later Solution at 5°C were equivalent to those stored at -80°C for at least four months after preservation.
The study concluded that RNA later Solution successfully preserved both RNA and DNA for up to one month at room temperature, and up to four months at 5°C, suggesting that RNA later Solution provides reliable sample preservation immediately upon collection. This result demonstrates that use of RNA later Solution for extended storage is equal to the established gold-standard of storage at -80°C. Also, with small organisms RNA later Solution provides the flexibility of storing them intact.
*nauplii is a developmental stage in brine shrimp.
RNAlater Tissue Collection: RNA Stabilization Solution
Total cellular RNA and DNA were quantified in individual nauplii using a fluorometric assay. RNA and DNA yields were subsequently compared with control samples (fresh animals assayed immediately after hatching) and among preservation treatments to determine the longterm effects of different sample storage techniques. Nucleic acid quantities were determined for each preservation treatment after one, two, four, and eight months. For both RNA and DNA yields, shrimp samples stored in RNA later Solution at 5°C were equivalent to those stored at -80°C for at least four months after preservation.
The study concluded that RNA later Solution successfully preserved both RNA and DNA for up to one month at room temperature, and up to four months at 5°C, suggesting that RNA later Solution provides reliable sample preservation immediately upon collection. This result demonstrates that use of RNA later Solution for extended storage is equal to the established gold-standard of storage at -80°C. Also, with small organisms RNA later Solution provides the flexibility of storing them intact.
*nauplii is a developmental stage in brine shrimp.
References
- Gorokhova E (2005) Effects of preservation and storage of microcrustaceans in RNAlater on RNA and DNA degradation, Limnol. Oceanogr Methods 3:143–148.