Mobio powermax dna isolation kit

Total DNA from the matrices were extracted using the Mobio PowerMax DNA Isolation Kit (MO BIO Laboratories, Inc., Carlsbad, CA) according to the manufacturer's instructions. DNA concentration was determined using a ND-1000 spectrophotometer (Nanodrop Technologies-ThermoFisher Scientific, Wilmington, DE). Up to 10, 000 ng DNA, in Tris-EDTA (10 mM Tris-HCl, 1 mM EDTA, pH 8.0)-cesium chloride (CsCl) solution, was put into Quick-Seal polyallomer tubes (13 mm × 51 mm, 5.1 mL; Beckman Coulter), adjusted to a buoyant density (BD) of ~1.77 g mL -1 , sealed and centrifuged at 178, 000 × g (20 o C), for 48 h using an ultracentrifuge (Beckman Optima XPN-100, Beckman Coulter, USA) (Song et al., 2015) (link). Then the centrifuged DNA was fractioned into 22 fractions. Each fraction of the DNA was purified using the Omega MicroElute DNA Clean-Up Kit (Omega Bio-Tek, Norcross, GA, USA).

Total genomic DNA was extracted from sediment and microbial mat samples using the MoBio Powermax DNA Isolation kit (MoBio Laboratories, USA) in accordance with the manufacturer's instructions. Approximately 1 -5 g (wet weight) of environmental material was used for the extractions, and two separate extractions were pooled for each sample. One of the two extractions replaced the bead-beating step with 30 minutes incubation at 65°C to mitigate extraction bias induced by both harsh and gentle extraction techniques.