Firestone Lab Group

Laboratory Protocols

Nitrogen and Carbon Assays

• KCl Extraction for NH₄⁺ and NO₃^-

• Nitrification Potential Assay

• Denitrifying Enzyme Assay (Denitrification Potential)

• ¹⁵N Pool Dilution

• Microbial Biomass Nitrogen and Carbon

• Persulfate Digestion for MBN and MB-15N

• Direct extraction (¹⁴C) chloroform fumigation

Microbiology

• How to snap-freeze cell cultures

• Extraction of bacterial DNA from soils

• Agarose gel protocol for separation of genomic DNA

• Isolation of petroleum degraders

• PLFA extraction

Instrumentation & Sample Preparation

• Lachat reagents and standards

• Startup Checklist for the HP5890

• Startup for O-I 1010 TIC-TOC analyzer

• Analysis of Polycyclic Hydrocarbons by FID (EPA Method 8100)

• Preparation of solid samples for ANCA-IRMS

• Diffusing KCl extracts for ¹⁵N

Molecular Methods

• Cloning

• Direct Counts

• Gel Analysis

• Genomic fingerprinting

• DNA Removal

• RNA Removal

• Soil DNA Extraction

• Soil RNA Extraction

• BDT Sequencing

• Big Dye

• EDTA/Ethanol Precipitation of Cycle Sequenced Products

• EDTA/NaOAc/Ethanol Precipitation of Cycle Sequenced Products

• 16S TRFLP for 96 Samples

• 16S TRFLP for Single Samples

• Phenol – Chloroform Purification

• Protease Assay

• Soil Fluorogenic Enzyme Assays

• Soil Moisture Content Measurement and Calculations

• Phosphate Buffered Saline (PBS)

• Phosphate buffer

• TE buffer

• Instructions for Rhizosphere Microcosms

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Miscellaneous

• Sterile hydroponics

• Chemical Inventory

Last updated: January 30, 2007