Illumina MiSeq paired end sequencing of SAMD00084889
The water of each sample was filtered through a 0.2 ????m membrane within 5 days of sampling. Each filter was quickly placed in a sterile tube and stored frozen at -20????C. DNA was extracted from the material on each filter using the PowerWater DNA Isolation Kit (MO BIO Laboratories, Carlsbad, CA) with bead homogenization at 3,000 rpm for 30 s. PCR amplification was performed using the primers 342F and 806R, which are V3-V4 region universal primers for prokaryotic 16S rRNA genes, and Ex Taq DNA polymerase (Takara, Shiga, Japan) with a denaturation step at 98????C for 2 min, followed by 30 cycles at 98????C for 10 s, 50????C for 30 s, and 72????C for 40 s, and a final extension step at 72????C for 10 min. PCR products were purified with the Agencourt AMPure XP system (Beckman Coulter).