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Antibody Addition and Detection for Staining Caenorhabditis elegans Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4523

Category: C. elegans Protocols: C. elegans Staining Protocols

Extreme care should be used to identify and verify positive reactions, however, because cross-reactions are common. Counterstaining is essential for examining worms by immunofluorescence and is used to identify the exact cell in which an antigen appears. Methods for counterstaining include labeling all cells with a fluorescent dye that is specific for nucleic acids (e.g., DAPI or propidium iodide) and using GFP driven by tissue-specific promoters. - [Read Antibody Addition and Detection for Staining Caenorhabditis elegans Protocol]

C. elegans Gene Knockout Protocol - http://www.zoology.ubc.ca/~alorch/koprotocol.htm

Category: C. elegans Protocols: C. elegans Genetics Protocols

The procedure is to mutagenize a large population of worms with trimethylpsoralen and UV irradiation, set up 1152 subpopulations, screen DNA made from this library for deletions in specific genes by nested PCR, and then to recover single worms carrying the deletions through a sib-selection process. - [Read C. elegans Gene Knockout Protocol]

C. elegans RNAi Protocol - http://www.zoology.ubc.ca/~alorch/rnai-protocol.htm

Category: C. elegans Protocols: C. elegans Genetics Protocols

Protocol for C. elegans RNAi. Includes: Transformation of competent cells; Blunt-end ligation; Preparation of competent cells; Dephosphorylation of linear plasmid DNA; Restriction Digest: EcoRV; Insert amplification from gDNA; Gel purification: QiaQuick gel purification kit; Mini-prep; Transformant Screening; Bacterial preparation and induction; Preparation of worms for RNAi feeding. - [Read C. elegans RNAi Protocol]

Cleaning Worms by Floating on Sucrose Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=18

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol for cleaning worms by floating on sucrose. - [Read Cleaning Worms by Floating on Sucrose Protocol]

Fixing Caenorhabditis elegans in Bouin’s Fixative Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4521

Category: C. elegans Protocols: C. elegans Fixing Protocols

Bouin’s fixative is a particularly good choice for worms because it penetrates dense tissues well and is extremely good for fixing antigens. Like all strong fixatives, however, it is unsuitable for some antibody-antigen pairs. In such cases, the length of time in the Bouin’s fixative can be shortened, or paraformaldehyde fixation can be used instead. - [Read Fixing Caenorhabditis elegans in Bouin’s Fixative Protocol]

Fixing Caenorhabditis elegans in Paraformaldehyde Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4522

Category: C. elegans Protocols: C. elegans Fixing Protocols

Common method for fixing worms is to use paraformaldehyde. This method provides a gentler fixation than the Bouin’s method, but often requires the use of collagenase. This method is particularly good for examining adult worms. - [Read Fixing Caenorhabditis elegans in Paraformaldehyde Protocol]

Freezing Worms Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=20

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol for freezing worms. Includes: The preferred method and The agar method. - [Read Freezing Worms Protocol]

Liquid Culture of Worms Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=22

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol for liquid culture of worms. Includes: superbroth; S- basal; worm plates; GROWING THE BACTERIA (WORM FOOD); GROWING THE WORMS; AFTER THE CULTURE HAS GROWN; PREPARING EGGS TO START SYNCHRONIZED LIQUID CULTURES. - [Read Liquid Culture of Worms Protocol]

Liquid Culture of Worms with Fermentor Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=21

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol for liquid culture of worms with fermentor. - [Read Liquid Culture of Worms with Fermentor Protocol]

Microinjecting C.elegans Worms Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=5

Category: C. elegans Protocols: C. elegans Injection Protocols

Protocol for microingecting C.elegans worms. Includes: Making the DNA solution; Setting up the scope, loading the needle, mounting, and breaking the needle; Mounting worms on an injection pad; Injecting. - [Read Microinjecting C.elegans Worms Protocol]

Phalloidin Staining of C. elegans Worms Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=8

Category: C. elegans Protocols: C. elegans Staining Protocols

Protocol for phalloidin staining of C. elegans worms. - [Read Phalloidin Staining of C. elegans Worms Protocol]

Preparation of Seeded NGM Plates For Worm Food Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1558

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol describes how to seed plates with E. coli OP50 for C. elegans cultures. Includes: Bacterial Stock Plate Preparation; Bacterial Broth Preparation; Seeding Bacteria Plates for Culturing Worms. - [Read Preparation of Seeded NGM Plates For Worm Food Protocol]

Preparation of Worm Extracts by Sonication Protocol - http://www.genetics.wustl.edu/tslab/Protocols/Cyto_extract_sonicat_v2.htm

Category: C. elegans Protocols

Protocol was designed to rapidly generate small scale cytosolic extracts of C. elegans for Western or IP (has not been tested for RNA work). The protocol works well for between 50 to 5000 worms and has not been extensively tested on larger a scale, though it should work. Includes: Collection; Sonication; Clearing lysate; Immunoprecipitation. - [Read Preparation of Worm Extracts by Sonication Protocol]

Pristionchus Pacificus Genetic Protocols - http://www.wormbook.org/chapters/www_ppageneticprotocols/ppageneticprotocols.pdf

Category: Developmental Biology: Utilizing Model Organisms Protocols

Protocols on the genetics of Pristionchus pacificus. Includes: Freezing worms; Mutagenesis; Construction of deletion libraries to generate P. pacificus gene knock-outs; RNAi and morpholino by injection. - [Read Pristionchus Pacificus Genetic Protocols]

Pristionchus pacificus Genetic Protocols - http://www.wormbook.org/chapters/www_ppageneticprotocols/ppageneticprotocols.pdf

Category: Laboratory Model Organisms Protocols

Genetic Protocols for Pristionchus pacificus. Includes: Freezing worms; EMS mutagenesis; Psoralen mutagenesis; Construction of deletion libraries to generate P. pacificus gene knock-outs; Designing primers for the gene of interest; RNAi and morpholino by injection. - [Read Pristionchus pacificus Genetic Protocols]

RUVKUN Antibody Staining Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=11

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for RUVKUN antibody staining. Includes: Fixation; To make solution; REDUCING DISULFIDES TO -SH; OXIDIZE -SH GROUPS TO -SO3; TO CHECK THAT WORMS ARE PERMEABLE TO MACROMOLECULES; ANTIBODY INCUBATIONS; PERMANENT SPRINGTIME MOUNTING. - [Read RUVKUN Antibody Staining Protocol]

RUVKUN Antibody Staining Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=11

Category: C. elegans Protocols: C. elegans Staining Protocols

Protocol for RUVKUN Antibody Staining. Includes: Fixation; To make solution; REDUCING DISULFIDES TO -SH; OXIDIZE -SH GROUPS TO -SO3; TO CHECK THAT WORMS ARE PERMEABLE TO MACROMOLECULES; ANTIBODY INCUBATIONS; PERMANENT SPRINGTIME MOUNTING. - [Read RUVKUN Antibody Staining Protocol]

Whole-Mount In Situ Hybridization for the Detection of RNA in C. elegans Embryos Protocol - http://cobweb.dartmouth.edu/~ambros/worms/14.html

Category: C. elegans Protocols

Protocol for whole-mount in situ hybridization for the detection of RNA in C. elegans embryos. Includes: Collection of Embryos; Permeabilization and fixation of embryos; Prehybridization; Hybridization; Post-hybridization washes; Probe Detection; Double-labeling; Interpretation. - [Read Whole-Mount In Situ Hybridization for the Detection of RNA in C. elegans Embryos Protocol]