title: Sewage Influent Bacterial SSU rRNA V6 tag sequencing, South Shore 8/21/07
design: A cocktail of five fused primers at the 5’ end of the V6 region (E. coli positions 967- 985) and four primers at the 3’ end (E. coli positions 1046-1028) captured the full diversity of rRNA sequences represented in molecular databases (Sogin et al., 2006; Huber et al., 2007). The 5’ fused primer includes a 5 nt “key” inserted between the Life Sciences primer A and the 967F rRNA primer.  We prepared amplicon libraries from at least three independent PCR cocktails for each sample to minimize the impact of potential early-round PCR errors, and sequenced using standard Roche protocols on a GS-FLX.