Metabarcoding of Ceratina_calcarata_EK2339_16S [Raw reads]
DNA extraction was performed using Omega E.Z.N.A. Soil DNA Kits, following the manufacturers protocol for 100-250mg samples, including bead lysis, with some modifications. To increase DNA yields the cHTR reagent recommended for soil extraction protocols was not used in this protocol, resulting in one less round of washing, centrifugation, and collection of the supernatant. Six blanks were included in all the DNA extraction steps and included in plate sample submission, library preparation, and sequence processing. Illumina MiSeq paired end amplicon sequencing was performed using the 16S rRNA region for bacteria and the ITS region for fungi. Specifically, the V5-V6 fragment was examined with the 799bF-CS1 forward primer (MGGATTAGATACCCKGG) and the 1115R-CS2 reverse primer (AGGGTTGCGCTCGTTG) for bacteria. The ITS1 fragment was used to examine fungi with the forward primer of ITS1F (CTTGGTCATTTAGAGGAAGTAA) and reverse primer of ITS2 (GCTGCGTTCTTCATCGATGC). Data was delivered as demultiplexed FASTQ files with adapters already trimmed. Library preparation and sequencing were performed by Genome Qubec Centre DExpertise et de Services.