Bacterial 16S rRNA amplicons from Lake Hazen: Summer 2015
The primers for each sequencing were used in a 28 cycle PCR with barcodes on the forward primer, using the HotStarTaq Plus Master Mix Kit (Qiagen, Germantown, MD, USA). Temperature cycling in PCR consisted of: 94 C, 3 min; 28 cycles of {94 C, 30 s; 53 C, 40 s; 72 C, 1 min} , with a final elongation at 72 C for 5 min. The PCR products were then checked in a 2% agarose gel and pooled in equal proportions based on the molecular weights and DNA concentrations of the products, followed by purification with calibrated Ampure XP beads (Beckman Coulter, CA, USA). All sequencing was performed on the Illumina MiSeq platform producing paired-end 300 bp reads according to the manufacturer s guidelines, by Molecular Research LP (Shallowater, TX, USA).
SUB3511250
hazensummer_27F