ena-STUDY-CHINESE ACADEMY OF FORESTRY-26-09-2018-10:23:40:737-583
Ectomycorrhizal fungi isolated with Quercus virginiana
From December 2017 to August 2018, ectomycorrhizal samples were collected from Quercus virginiana forest in Shangyu, Zhejiang four times. Genomic DNA from ectomycorrhizal root samples was extracted using CTAB method. We utilized internal transcribed spacer 2 (ITS2) as a barcode for pyrosequencing, which was amplified using fITS7 (5'-GTGARTCATCGAATCTTTG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC- 3') primer pair (Ihrmark et al. 2012). All PCR reactions were carried out with Phusion® High– Fidelity PCR Master Mix (New England Biolabs). Then, samples were purified with Qiagen Gel Extraction Kit (Qiagen, Germany). Library preparation and sequencing Library was generated using TruSeq® DNA PCR–Free Sample Preparation Kit (Illumina, USA), then its quality was assessed on the Qubit@ 2.0 Fluorometer (Thermo Scientific) and Agilent Bioanalyzer 2100 system. The amplicon library was sequenced on the Illumina HiSeq2500 platform and 250 bp paired–end reads were generated.
From December 2017 to August 2018, ectomycorrhizal samples were collected from Quercus virginiana forest in Shangyu, Zhejiang four times. Genomic DNA from ectomycorrhizal root samples was extracted using CTAB method. We utilized internal transcribed spacer 2 (ITS2) as a barcode for pyrosequencing, which was amplified using fITS7 (5'-GTGARTCATCGAATCTTTG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC- 3') primer pair (Ihrmark et al. 2012). All PCR reactions were carried out with Phusion® High– Fidelity PCR Master Mix (New England Biolabs). Then, samples were purified with Qiagen Gel Extraction Kit (Qiagen, Germany). Library preparation and sequencing Library was generated using TruSeq® DNA PCR–Free Sample Preparation Kit (Illumina, USA), then its quality was assessed on the Qubit@ 2.0 Fluorometer (Thermo Scientific) and Agilent Bioanalyzer 2100 system. The amplicon library was sequenced on the Illumina HiSeq2500 platform and 250 bp paired–end reads were generated.