Taxus_contorta_root_AMfungal_metabaroding
Amplification of the targeted V4 region of the 18S rRNA was carried out. Primers used were nu-SSU-0595-5' CGGTAATTCCAGCTCCAATAG and nu-SSU-0948-3' TTGATTAATGAAAACATCCTTGGC (Dagher et al. 2020).The QC passed amplicons with the Illumina adaptor were amplified using i5 and i7 primers that add multiplexing index sequences as well as common adapters required for cluster generation (P5 and P7) as per the standard Illumina protocol. The amplicon libraries were purified by AMPureXP beads and quantified using Qubit fluorometer. The amplified library was analyzed on 4200 Tape Station system (Agilent Technologies) using D1000 Screen tape as per manufacturer instructions. After obtaining the mean peak size from Tape Station profile, libraries were loaded onto MiSeq at appropriate concentration (10pM) for cluster generation and sequencing.