Global biogeography of desert cyanobacteria

Study Extent

Sampling

Method steps

1. Amplification of 16S rRNA genes was achieved using primer pair 341F and 907R with PCR conditions involving an initial denaturation time of 5min; 30 cycles at 95°C for 1 min, 55°C for 1 min, 72°C for 1 min, and a final extension at 72°C for 10 min. Positive and negative controls were run for every PCR. For each amplicon library purification was carried out with Agencourt AMPure XP Bead (Beckman Coulter, CA, USA) according to manufacturer’s instructions. The libraries were quantified with Quant-iT PicoGreen dsDNA Assay Kit (Invitrogen Life Technologies, NY, USA) using FLUOstar OPTIMA F fluorometer (BMG Labtech GmbH, Offenburg, Germany) and library quality was assessed with the FlashGel System (Lonza Group Ltd., Basel, Switzerland).
2. Emulsion-PCR was carried out with GS Junior Titanium emPCR Kit (Lib-L, 454 Life Sciences Corp., CT, USA) according to the emPCR Amplification Method Manual – Lib-L, Single-Prep. The sequencing reaction was carried out with the GS Junior Titanium Sequencing Kit and GS Junior Titanium PicoTiterPlate Kit (454 Life Sciences Corp.) according to the manufacturer’s instructions. The sequencing run was conducted in 200 cycles.

1. Bacteria
   common name: Bacteria rank: domain
2. Cyanobacteria
   common name: Cyanobacteria rank: phylum