Model 7670541 freezone 2.5 liter

Fresh samples were weighed and frozen for two days at −80 °C. Then, the samples were taken to a freeze–evaporation system (Model 7670541 FreeZone 2.5 Liter Labconco Freeze Dry Systems) and all the water contained in the original product was removed by freeze–evaporation cycles. A total of 3 g of each dried lichen was macerated with methanol (3 times, 30 mL each time, 3 days/extraction). The solutions were concentrated to obtain 11 mg of extract from S. pseudosylvatica; 9 mg S. luteocyphellata; 14 mg S. cf. andina; 12 mg S. cf. hypoglabra; 10 mg S. cordillerana; 9 mg S. cf. gyalocarpa; 13 mg S. leucoblepharis; 13 mg S. parahumboldtii; 8 mg S. impressula and 9 mg S. ocaniensis, respectively. Then, the lichen extracts were processed individually for HPLC-MS analyses (redissolved in methanol at a concentration of 1 mg/mL for the analyses).

The biomass of the haloarchaea strains was obtained by centrifugation and stored at −80 °C for two days. Then, the samples were lyophilized in a freeze-evaporation system (Model 7670541 FreeZone 2.5 Liter Labconco Freeze Dry Systems, Palo Alto, CA, USA) where all water contained in the original product was removed by freeze-evaporation cycles. The lyophilizate was stored in 15 mL Falcon centrifuge tubes protected from light, and once the dry weight was determined, it was stored at −20 °C until use. The lyophilized material (0.3–1.0 g) was added 2 mL of an acetone: water mixture in a proportion (8: 2 v/v), [25 (link)], vortexed for 10 min, then sonicated for 10 min and subsequently centrifuged at 4500 rpm for 15 min at −4 °C. Three cycles of vortexing, ultra-sounding and centrifugation were employed until a final volume of 5 mL of supernatant was recovered, which was then stored at −20 °C protected from light until use.