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A Targeted Screen to Detect Recessive Mutations that have Quantitative Effects Protocol - http://www.nervenet.org/papers/Consomic.html

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

Describes an experimental cross in mice that can be used to define and map induced germ-line mutations that map to a single chromosome. The cross is a modification and extension of a conventional three-generation recessive mutagenesis screen. Includes: The Mutagenesis Breeding Plan; Consomic Strains; Generating Mutations; Generating and Genotyping G2 Females; Genotyping G3 Progeny; Phenotyping G4 Progeny; etc.. - [Read A Targeted Screen to Detect Recessive Mutations that have Quantitative Effects Protocol]

Confocal Microscopy Specimen Preparation Using Synthetic Fluorophores and Immunofluorescence - http://www.olympusconfocal.com/applications/protocols/index.html

Category: Microscopy Protocols: Confocal Microscopy Protocols

Includes staining protocols: Triple-Staining Adherent Cells with MitoTracker, Phalloidin,& Nuclear Dyes; Staining Adherent Cells with Cytokeratin Primary Antibodies & Synthetic Fluorophores; Staining Adherent Cells with Intermediate Filament Primary Antibodies & Synthetic Fluorophores; Staining Cells & Tissue Cryosections with Tubulin Primary Antibodies, Phallotoxins,& Synthetic Fluorophores; Triple-Staining Tissue Cryosections with Wheat Germ Agglutinin, Phalloidin,& Nuclear Dyes; etc. - [Read Confocal Microscopy Specimen Preparation Using Synthetic Fluorophores and Immunofluorescence]

Germ Layer Explant Recombination Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4369

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

This protocol describes a method for recombining and culturing germ layer fragments. It is useful for testing the inductive properties of fragments from wild-type and mutant mouse embryos. - [Read Germ Layer Explant Recombination Culture Protocol]

Germ Layer Explant Recombination Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4369

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol describes a method for recombining and culturing germ layer fragments. It is useful for testing the inductive properties of fragments from wild-type and mutant mouse embryos. - [Read Germ Layer Explant Recombination Culture Protocol]

Human Embryonic Stem Cell Protocols - http://stemcells.nih.gov/staticresources/research/protocols/BresaGen_hESC_manual_2.1.pdf

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Human embryonic stem cells (hESCs) are pluripotent cells capable of differentiation to representatives of all three germ layers. Includes: Isolation of Primary Mouse Embryo Fibroblasts; Thawing and preparing p1 MEF feeder plates; Preparation of MEF- Conditioned Medium (MEF-CM; Microdissection Passaging of hESCs; Bulk passaging of hESC; Cryopreservation of hESCs; Thawing of hESCs; Karyotyping. - [Read Human Embryonic Stem Cell Protocols]

Isolating Germ Cells from the Genital Ridge - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4370

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol describes isolation of germ cells from the genital ridge of fetal mice from 11.5 days post coitum (dpc) onward. The germ cells can then be used for analysis, culture, or transplantation. - [Read Isolating Germ Cells from the Genital Ridge]

Isolating Germ Cells from the Genital Ridge Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4370

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

This protocol describes isolation of germ cells from the genital ridge of fetal mice from 11.5 days post coitum (dpc) onward. The germ cells can then be used for analysis, culture, or transplantation. - [Read Isolating Germ Cells from the Genital Ridge Protocol]

Lectin-Agarose Affinity Chromatography Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4202

Category: Chromatography Protocols: Affinity Chromatography Protocols

Two methods are provided for purifying glycoproteins using wheat-germ agglutinin or concanavalin A-Sepharose. Because lectin-affinity matrices can bind a few milligrams of protein per milliliter of affinity matrix, only a small amount of affinity gel matrix is required. The batchwise method is recommended when protein volume is large. - [Read Lectin-Agarose Affinity Chromatography Protocol]

Mouse Gonad Removal and Germ Cell Staining Protocol - http://www.swarthmore.edu/NatSci/sgilber1/DB_lab/Mouse/mouse_gonads.html

Category: Mouse Protocols: Mouse Reproductive System Surgery Protocols

Protocol describes how to remove the gonads, determine the sex ofmouse embryos 13.5 days post coitum and visualize the germ cells, through use of an alkaline phosphatase staining. - [Read Mouse Gonad Removal and Germ Cell Staining Protocol]

Separating Postimplantation Germ Layers Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4368

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Germ layers are multipotent tissues that have the ability to differentiate into various tissue types. Isolation and manipulation of germ layers is important for examining differentiation. This protocol describes a method for separating postimplantation germ layers. - [Read Separating Postimplantation Germ Layers Protocol]

X-Ray Mutagenesis of Drosophila Protocol - http://www.bioprotocol.com/protocolstools/protocol.jhtml%3Bjsessionid%3DGYZOPNVU3IQQTR3FQLMSFEWHUWBNQIV0?id=p32

Category: Drosophila Protocols: Drosophila Genetics Protocols

Protocol describes the general procedure for creating mutations in the DNA of Drosophila by exposure to X-rays. Irradiation of cells with X-rays creates double strand breaks (DSBs) in DNA. Mutations introduced in the DNA of germ line cells (sperm) are propagated by mating the exposed males to virgin females. The progeny of this cross can be mated to each other so that a percentage of the subsequent offspring will have two copies of the same mutant allele. - [Read X-Ray Mutagenesis of Drosophila Protocol]

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Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.