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Protocols

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9 protocols

analyticalstandardized

DNA metabarcoding

Surface sterilization of plant tissues followed by DNA extraction and high-throughput sequencing of the ITS region to characterize fungal endophyte communities. Community composition and diversity met...

molecular20 papersplastic resealable bags, −20°C freezer...
analyticalstandardized

Illumina paired-end sequencing

Standard Illumina library preparation using Nextera XT kit followed by paired-end sequencing on HiSeq 2500 platform to generate comprehensive metagenomic datasets from phylloplane microbial communitie...

molecular4 papersNextera XT kit, SPRI beads...
analyticalstandardized

U.S. Geological Survey sediment preparation procedures

Extraction of iron and manganese from sediments using 0.5 N HCl with hydroxylamine hydrochloride in anaerobic conditions, followed by ICP-AES analysis. This method targets metals associated with reduc...

geochemical extraction3 papersHDPE bottles, 0.5 N HCl...
analyticalstandardized

FTIR with ATR accessory (Pinaceae)

Fourier Transform Infrared spectroscopy analysis of ground needle samples to quantify functional group concentrations including aromatics, ethers, amides, and carbonyls. Uses attenuated-total-reflecti...

molecular2 papersNicolet iS50 FTIR spectrometer, attenuated-total-reflection accessory
analytical

Manual curation of large extrachromosomal elements (Archaea)

Manual identification and curation of large genome fragments using GC content, coverage patterns, and taxonomic profiles to assemble complete extrachromosomal genomes.

genome assembly2 papersIDBA-UD, MEGAHIT...
analyticalstandardized

Metagenomic HMM metabolic profiling

Comprehensive Hidden Markov Model analysis of metagenomic sequences to identify and quantify metabolic functions across biogeochemical pathways. Uses both standardized TIGR-FAM and custom HMM database...

molecular2 papersTIGR-FAM database, Custom HMM databases
analyticalstandardized

maximum likelihood phylogeny

Construction of phylogenetic trees using concatenated ribosomal protein genes to determine evolutionary relationships. Uses IQ-TREE with maximum likelihood approach and bootstrap support.

phylogenetic2 papersIQ-TREE software
analytical

Anoxic TEM-EDS elemental mapping

Transmission electron microscopy with energy dispersive X-ray spectroscopy performed under anoxic conditions using vacuum transfer holders to map elemental distributions in redox-sensitive particles.

microscopy1 paperlacey carbon Cu grids, Fischione vacuum transfer holder Model 2560...
analytical

LC-MS plant metabolomics (Poaceae)

Extraction of plant metabolites using methanol/acetonitrile/water solvent followed by liquid chromatography-mass spectrometry analysis. Characterizes plant metabolomic profiles for correlation with fu...

metabolomics1 paperHPLC grade solvents, Synergi 2.5 μm reverse-phase Hydro-RP column...