title: ITS rRNA (reverse transcribed to cDNA) metabarcoding of soil fungi: 30-37.5 cm below soil surface beneath a dead Quercus douglasii
design: RNA was extracted from flash-frozen soil samples. RNA was purified and reverse transcribed to cDNA. The ITS region was amplified via PCR using targeted primers ITS4_Fun and 5.8S_Fun. 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.