title: 16S rRNA (reverse transcribed to cDNA) metabarcoding of soil bacteria: 0-7.5 cm below soil surface beneath a living Quercus douglasii
design: RNA was extracted from flash-frozen soil samples. RNA was purified and reverse transcribed to cDNA. The 16S region was amplified via PCR using targeted primers 515F and 806R. Sample-specific barcodes (synthetic oligonucleotides) were added to all initially amplified DNA molecules in a second round of PCR. All PCR products from this study were pooled, sequenced together, and demultiplexed based on barcode sequences.