16S rRNA genes of bacterioplankton community: a sample from a microcosm study
After the 10-day incubation, 500-mL water samples from each device were filtered using 0.2-m membrane filters (Millipore, Merck), which filters were then transferred into autoclaved cryopreservation tubes and fully immersed by adding 1 mL Ambion RNAlater to the filters. The entire cryopreservation tube was rapidly frozen in liquid nitrogen. After complete freezing, the tubes were transferred to a refrigerator at -80 for storage until RNA was extracted. The V4V5 region of the 16S rRNA gene was amplified by PCR using the primers 515F/926R (Parada et al. 2016).