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Assembling Aggregates between Diploid Embryos Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4423

Category: Mouse Protocols: Assembling Chimeras Mouse Protocols

Protocol describes a method for assembling aggregates between diploid embryos. If embryos from a heterozygous mutant intercross are aggregated with wild-type embryos, the resulting chimeras can be used for analyzing mutant phenotypes. - [Read Assembling Aggregates between Diploid Embryos Protocol]

Detection of Mutations by Single-strand Conformational Polymorphism Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3815

Category: Genetics and Genomics: Cancer Genetics Protocols: Single Strand Conformational Polymorphism(SSCP) Protocols

Single-strand conformational polymorphism (SSCP), one of several methods used to scan segments of DNA for mutations, exploits the electrophoretic differences in mobilities between single-stranded mutant and wild-type DNAs. - [Read Detection of Mutations by Single-strand Conformational Polymorphism Protocol]

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]

How to Choose Wild Type Laboratory Strains of N. crassa for Reference and for Special Uses - http://www.fgsc.net/neurosporaprotocols/How%20to%20choose%20wild%20types%20for%20reference%20and%20special%20uses.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to choose wild type laboratory strains of N. crassa for reference and for special uses. - [Read How to Choose Wild Type Laboratory Strains of N. crassa for Reference and for Special Uses]

How to Convert Wild-Type Spreading Growth to Colonial - http://www.fgsc.net/neurosporaprotocols/How%20to%20convert%20wild-type%20spreading%20growth%20to%20colonial.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to convert wild-type spreading growth to colonial. - [Read How to Convert Wild-Type Spreading Growth to Colonial]

How to Determine the Species of a Wild-Collected Isolate - http://www.fgsc.net/neurosporaprotocols/How%20to%20determine%20the%20species%20of%20a%20wild-collected%20isolate.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to determine the species of a wild-collected isolate. - [Read How to Determine the Species of a Wild-Collected Isolate]

Protocol Live Imaging of Caenorhabditis Elegans - http://www.cshprotocols.org/cgi/content/extract/2006/29/pdb.ip19

Category: Microscopy Protocols: In Vivo Imaging Protocols

To image early cleavages and chromatin dynamics, it is convenient to use histone H2B fused to GFP or lamin::GFP. Time-lapse movies can be obtained using conventional confocal microscope systems and their included software. Early embryos dissected from transgenic hermaphrodites are placed with egg salts on agar pads. Chromatin dynamics can be followed easily, and wild-type embryonic cells can be compared with mutants or RNAi-treated embryos. - [Read Protocol Live Imaging of Caenorhabditis Elegans]

Screening Recombinant Clones for Site-directed Mutagenesis by Hybridization to Radiolabeled Oligos - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4013

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Plaques formed by M13 bacteriophages or bacterial colonies transformed by plasmids carrying specific mutations can be detected by hybridization, using a radiolabeled oligonucleotide that forms a perfect duplex with the mutant sequence. Hybridization is carried out under conditions of low stringency that allow the radiolabeled oligonucleotide to anneal to both mutant and wild-type DNAs. - [Read Screening Recombinant Clones for Site-directed Mutagenesis by Hybridization to Radiolabeled Oligos]

Single tube confirmation PCR protocol - http://www-sequence.stanford.edu/group/yeast_deletion_project/single_tube_protocol.html

Category: PCR Protocols: Colony PCR

The following colony PCR protocol has been designed to be performed in individual reaction tubes. We usually test three colonies from each transformation along with a wild-type control. A series of five different PCR tests are performed on each colony to - [Read Single tube confirmation PCR protocol]

Transfection of Bone Marrow-Derived Mast Cells for Transcription Factor Luciferase Reporter Assays - http://www.natureprotocols.com/2006/09/12/transfection_of_bone_marrowder_1.php

Category: Reporter Gene Assays: Luciferase Assay Protocols

Transfection of primary leukocytes has traditionally been a challenging but much desired protocol. It allows not only the analysis of cells in a more natural state to a cell line system, it enables the direct comparison of, for e.g. transcriptional activity using luciferase reporters, in immune cells taken from genetically-altered mice. In addition, importantly it allows for "rescue experiments" in knockout cells & the ability to over-express or reconstitute wild-type and/or mutated constructs. - [Read Transfection of Bone Marrow-Derived Mast Cells for Transcription Factor Luciferase Reporter Assays]

Articles (1-1 of 1)

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.