Mycelia of the fungal isolates were grown in flasks with half-concentration potato dextrose broth (PDB) liquid medium for 3 to 7 days (depending on fungal growth) and collected in 2-ml tubes for DNA extraction. Total DNA was extracted with the NucleoSpin Plant II kit (MACHEREY–NAGEL, Düren, Germany) according to the manufacturer’s instructions. The quality and quantity of DNA samples were assessed by spectrophotometry (ND-1000 Spectrophotometer NanoDropH; Thermo Scientific, Germany). The nuclear ribosomal ITS region was amplified from all DNA extracts with the ITS1-ITS4 fungal primers (White et al. 1990 ). The ITS representative sequences of OM isolates generated in this study were submitted to GenBank and recorded under the following string of accession numbers: OM971071-OM971074 for Ceratobasidiaceae and OM976746-OM976769 for Tulasnellaceae.
Nd 1000 spectrophotometer nanodroph
Manufactured by Thermo Fisher Scientific
Sourced in Germany
The ND-1000 Spectrophotometer NanoDropH is a compact and versatile laboratory instrument designed for the quantification and analysis of DNA, RNA, and protein samples. The device utilizes a unique sample-retention system that requires only 1-2 microliters of sample to perform measurements, minimizing sample consumption. The NanoDropH provides accurate and reproducible results, making it a valuable tool for researchers and scientists working in various fields, including molecular biology, genetics, and biochemistry.
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2 protocols using nd 1000 spectrophotometer nanodroph
1
Fungal DNA Extraction and Sequencing
2
Genome-skimming for microsatellite detection
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Total genomic DNA was extracted from silica-dried leaf tissues using the DNeasy Plant Mini Kit (QIAGEN) according to the manufacturer's instructions, after disruption with TissueLyser. The quantity and quality of DNA samples were assessed by spectrophotometry (ND-1000 Spectrophotometer NanoDropH; Thermo Scientific, Wilmington, Germany) and by electrophoresis on a 1% agarose gel.
A genome-skimming approach (Straub et al. 2012; Dodsworth 2015) was used for the detection of microsatellites (Viruel et al. 2018; (link)Landoni et al. 2020) . Genomic libraries were prepared using NEBNext® UltraTM II DNA Library Prep Kit for Illumina® (New England Biolabs, Ipswich, MA, United States) with AMPure XP magnetic beads for size selection (300-350 bp) and NEBNext® Multiplex Oligos for Illumina® (Dual Index Primer Sets I and II) as barcodes for simultaneous sequencing (Viruel et al. 2019) (link). Library quality was evaluated using a QuantusTM fluorometer (Promega Corp.) and an Agilent 4200 TapeStation (Agilent Technologies, Santa Clara, CA, United States). Multiplexed libraries were then sequenced on an Illumina MiSeq (Illumina, Inc.) lane. Trimmomatic v0.35 (Bolger et al. 2014 (link)) was used to remove low-quality reads and adapter sequences flagged by FastQC v0.11.7 (Andrews 2010) .
A genome-skimming approach (Straub et al. 2012; Dodsworth 2015) was used for the detection of microsatellites (Viruel et al. 2018; (link)Landoni et al. 2020) . Genomic libraries were prepared using NEBNext® UltraTM II DNA Library Prep Kit for Illumina® (New England Biolabs, Ipswich, MA, United States) with AMPure XP magnetic beads for size selection (300-350 bp) and NEBNext® Multiplex Oligos for Illumina® (Dual Index Primer Sets I and II) as barcodes for simultaneous sequencing (Viruel et al. 2019) (link). Library quality was evaluated using a QuantusTM fluorometer (Promega Corp.) and an Agilent 4200 TapeStation (Agilent Technologies, Santa Clara, CA, United States). Multiplexed libraries were then sequenced on an Illumina MiSeq (Illumina, Inc.) lane. Trimmomatic v0.35 (Bolger et al. 2014 (link)) was used to remove low-quality reads and adapter sequences flagged by FastQC v0.11.7 (Andrews 2010) .
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