454 pyrosequencing for microbial community study
DNA extracts were amplified with: barcode specific primers ITS1F/ITS2 that amplify the ITS1 hypevariable region of the ITS rRNA. The forward primer od each primer pair used holds a unique 10-base sample identification barcoding tags to separate samples after sequencing. Three PCR reactions were carried out for each sample and then pooled. PCR products were purified and quantified with Bioanalyser. Equivalent molarity of each of the amplicon-pools were bulked for the subsequent pyrosequencing with the 454 Life Sciences GS FLX system.