Note: teaching protocols and materials can be found at "For Teachers: Teaching materials by Fields of Study" under C. elegans education.

Commonly used C. elegans Research protocols

Worm Culture


• Dauer Pheromone Prep
• Making dauers with pheromone


• Bacteria for worm food
• “Chunking” worms- easy strain maintenance
• Cleaning worms by floating on sucrose
• Cleaning worm stocks
• Cleaning Worm Stocks
• Cloning form low melt agarose
• Cool growth competent E. coli
• Culturing Worms
• Decontaminating worm population to remove fungi and bacteria
• Development of techniques for primary culture of C. elegans
• Development of techniques for primary culture of C. elegans embryonic neurons Laird Bloom
Part 1
• Development of techniques for primary culture of C. elegans embryonic neurons
Part 2
Part 3
• ES cell culture
• Freezing worms
• Growing worms in liquid
• Liquid culture of worms #1
• Liquid culture of worms #2
• Liquid culture of worms in with fermentor
• Maintenance of lin2 strain C.elegans Worms
• N2 Development time at different temperatures #1
• N2 development times at different temperatures #2
• Preparation pf Petri plates
• Preparation of synchronized cultures (PDF)
• Preparation of Seeded NGM Plates For Worm Food
• Response to food
• Quick bleach cleaning of small numbers of worms
• Quick bleach decontamination of small numbers of C. elegans
• Synchronizing worm cultures
• Testing for Levasimole resistance




• Buffers
• Embryo buffers


• S-medium, S-basal and trace metal solution



• C. elegans gene knockout protocol
• Chemical deletion mutagenesis
• DpnI mediated site-directed mutagenesis
• EMS mutagenesis
• ems mutagenesis
• Gene knockout with conventional mutagenesis
• Irradiation of worms in Y-building facility
• Making males by heat shock
• Transposon Insertion Mutagenesis


• Mapping mutants using STS Polymorphisms
• Snip-SNP mapping with BSA
• Snip-SNP mapping with Polymorphisms
• SNP characteristics


• F2 mutant screen
• Oligos for screening of deletion libraries
• Rapid screen of transformants



• Creating Transgenic C. elegans with Extrachromosomal Arrays
• Integrating extrachromosonal arrays
• Microinjecting worms
• Procedures for preparing pads and needles
• Transformation by injection into syncytial C.elegans germline


Molecular & Biochemical Techniques


• Western Blot Transfer and Detection
• Worm genomic southern blots
• Worm extracts and western blotting

Cell Culture

• Development of techniques form primary culture of embryonic neurons


• cDNA amplification
• Gel purification of DNA fragments
• Genomic DNA preparation
• Phenol extraction from low melt agarose
• Plasmid DNA prep
• Silica clean-up of DNA
• Worm genomic DNA prep
• “In gel-o” ligations
• Cosmid Preparation
• Cosmid Vectors Codes for C. elegans


• aRNA amplification
• Joe’s mRNA preparation
• Nematode genomic DNA isolation
• RNA isolation (TRIZO method)
• RNA preparation
• RNA synthesis (in vitro transcription)
• Total RNA preparation
• Yale RNA preparation


• PCR analysis of dead eggs from C. elegans
• Polymerase Chain Reaction
• Single Worm PCR #1
• Single Worm PCR #2
• Single egg PCR reactions


• Affinity Purification of Antibodies to GST-fusion Proteins
• Bacillus thuringiensis (Bt) Toxin Purification
• Electrotransfer of proteins (electroblotting)
• Immunoprecipation
• Preparation of total SDS-soluble nematode proteins
• Preparation of GST-capping protein
• Protease inhibitors
• Purification of MBP Fusion Protein
• Purification of GST-Fusion Protein
• SDS- Page

Transforming Worms

• Integrating extrachromosanal array into C. elegans chromomosomes
• Large Scale Transformation by microparticle bombardments



• 96-well format RNAi in liquid culture
• Bacteria mediated RNAi protocol
• dsRNA preparation for RNAi by injection in C. elegans (PDF)
• Making RNAi primers
• RNAi experiments
• Double Stranded RNA interference (RNAi ) in C. elegans
• Microinjection of RNA into C. elegans
• RNA injection



Antibody staining

• Antibody purification and Testing
• C. elegans P-granule antibody staining
• C. elegans Hermaphrodite or Embryo Antibody Staining: Seydoux and Dunn Method
• DCF staining
• Freeze- fracture for Immunofluorescence of Embryos
• Immunofluorescence of C. elegans Embryos
• Serotonin Antibody Straining
Short version
Long and detailed version
• Solutions for Anti-Serotonin protocol
• PAR1-staining
• Preabsorbing Antisera with C. elegans Acetone Powder


• Electron Microscopy (EM) Fixation of C. elegans Embryos
• EM fixation of embryos
• Fixation
• Fixation and Permeabilization
• Fixation and staining parameters
• Standard Osmium/Glutheraldehyde Fixation
• Microwave fixes


• Bacillus Thuringiensis Crystal Staining
• C. elegans: Whole Animal DAPI Staining
• Double dye filling
• Dye filling to stain amphid and phasmid neurons
• In situ hybridization
• Gonad dissections, fixation; DAPI staining, antibody staining, germline in situ hybridization
• Mms integration
• Nematode dye filling
• Phalloidin Staining
• RNA in situ hybridization of dissected gonads
• Staining C. elegans for B-galactosidase



• Electron Microscopy (EM) Fixation of C. elegans Embryos
• EM fixation of embryos
• Fluorescent dissecting stereo microscope
• Laser Ablation System

Other techniques

• Fermentor
• Gonad dissections
• Lethal phase determination
• Mms integration
• Quick and easy yeast transformation
• The Electropharyngeogram