ITSKG07
Fungal Community diversity
Next generation sequencing library preparations and Illumina MiSeq sequencing were conducted.DNA samples were quantified using a Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA, USA). 50-100ng DNA was used to generate amplicons using a panel of primers designed.Oligonucleotide primers were designed to anneal to the relatively conserved sequences spanningfungi ITS regions. ITS2 region was amplified using forward primer containing sequence"GTGAATCATCGARTC" and reverse primer containing sequence "TCCTCCGCTTATTGAT".Besides the ITS target-specific sequences, the primers also contain adaptor sequences allowinguniform amplification of the library with high complexity ready for downstream NGS sequencing onIllumina Miseq platform.DNA libraries were validated by Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA,USA), and quantified by Qubit 2.0 Fluorometer. DNA libraries were multiplexed and loaded on anIllumina MiSeq instrument according to manufacturers instructions (Illumina, San Diego, CA,USA). Sequencing was performed using a 2x300/250 paired-end (PE) configuration; image analysisand base calling were conducted by the MiSeq Control Software (MCS) embedded in the MiSeqinstrument.
ITSKG07
SUB4273777
ITSKG07