show Abstracthide AbstractIn order to increase the depth of sampling for diverse bacterial communities, we developed a new method for sequencing and assembling millions of paired-end reads from the 16S rRNA gene (spanning the V3 region; ~200 nucleotides) using Illumina sequencing platforms. Sequence libraries were prepared in duplicate for both a defined mixture of DNA from known bacterial cultured isolates (>1 million sequences combined) and from an arctic tundra soil sample (>6 million sequences following initial filtering).