TABLE 2.

Primers used for qPCR assays, annealing temperatures, target regions, and the specificity of the amplicons cloned from qPCR assays with soil DNAa

Target groupForward primerReverse primerApproximate amplicon length (bp)Annealing temp (°C)% of soil clones belonging to the target groupc
All BacteriaEub338Eub51820053100
α-ProteobacteriaEub338Alf6853656075
β-ProteobacteriaEub338Bet6803606096
Actinobacteria Actino235Eub5183006060
Firmicutes Lgc353Eub51818060100
Bacteroidetes Cfb319Eub51822065100
Acidobacteria Acid31Eub51850050100
All Fungi5.8sITS1f300b53100
Basidiomycota ITS4b5.8sr500b55100
  • a The bacterial qPCR assays target 16S rRNA genes, while the fungal assays target the internal transcribed spacer region found in rRNA genes.

  • b The length of the targeted ITS region can vary significantly between different fungal strains.

  • c The percentage includes only those clones that were nonchimeric and exceeded the 80% confidence threshold for taxonomic assignment using the RDP classifier program.