transgenic Protocols

Category: Cytogenetics Protocols

Protocol for Agrobacterium-mediated transformation in rice. Agrobacterium-mediated rice transformation method that is applicable to easily cultured varieties in addition to elite japonica varieties that are more difficult to culture. Using this method, transgenic rice plants can be obtained in about 2–3 months with a transformation frequency of 30–50%, both in easily cultured varieties and recalcitrant elite japonica rice. - [Read Agrobacterium-Mediated Transformation in Rice Protocol]

Category: Immunology: T Cell Protocols

Protocol for culture and expansion of OT-1 TCR transgenic T cells. - [Read Culture and Expansion of OT-1 TCR Transgenic T cells Protocol]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Mouse Embryonic Stem Cell Culture. Southern Analysis of ES cells. The Darwin Transgenic Core Facility - [Read ES mouse cell stem cell Culture]

Category: Transgenic Mouse Protocols: Mouse Genotyping Protocols

Test mice and rats for the presence of the transgene by PCR. University of Michigan, Transgenic Animal Model Core - [Read Genotyping Transgenic Rodents by PCR]

Category: Cytoskeleton Protocols: Actin Myosin Protocols

The same GFP-tagged actin construct used in cell transfection experiments has been used to produce transgenic mice. Transgenic animals allow the imaging of brain tissue in the intact animal, as acutely cut slices or as organotypic slice cultures. They also serve as a source of cells for imaging neurons at high resolution in dispersed low-density cell culture. In contrast to cells transfected in culture, where the level of actin-GFP expression in neurons varies considerably, transgenic mice... - [Read Imaging Actin in Tissue Slices from Transgenic Mouse Brain Protocol]

Category: Transgenic Mouse Protocols: Mouse Tail DNA Extraction Genotyping

Isolation of DNA from Mouse Tail Biopsies. University of Michigan Transgenic Animal Model Core, Transgenic Animal Web. - [Read Isolation of DNA from Mouse Tail Biopsies]

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

The most commonly used markers for selection of transgenic Arabidopsis are resistance to the antibiotic kanamycin and to the herbicide glufosinate ammonium. Resistance to kanamycin is conferred by a bacterial gene encoding the enzyme neomycin phosphotransferase (NPT). In this protocol, kanamycin-resistant seedlings are selected on solid medium. - [Read Kanamycin Selection of Transformed Arabidopsis Protocol]

Category: Yeast Protocols: Yeast Genetics Protocols: YACs Protocols

Protocol describes the use of agarose plugs for isolation of yeast artificial chromosome (YAC) DNA. The DNA can then be run on a pulsed-field gel and used for microinjection to produce transgenic mice. - [Read Large-Scale Preparation of Agarose Plugs of Yeast DNA Protocol]

Category: C. elegans 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. - [Read Live Imaging of Caenorhabditis elegans: Examples]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Mammalian toxicology repeat dose toxicity studies. Includes: M204 14-Day Oral Toxicity Study (Rodents), M204-H, M204-RH, M205 14-Day Oral Toxicity Study (Dogs), M205-H, M205-RH, M201 28-Day Oral Toxicity Study, M201-R, M202 28-Day Oral Toxicity Study, M202-R, M211 90-Day Oral Toxicity Study (Rodent), M211-20, M212 90-Day Oral Toxicity Study (Dog), M215 90-Day Dermal Toxicity Study, M213 Two-Year Chronic Toxicity/Carcinogenicity Study, M214 6-Month Carcinogenicity Study in p53+/Transgenic Mice. - [Read Mammalian Toxicology Repeat-Dose Toxicity Studies]

Category: Microinjection Protocols: Microinjection of DNA Protocols

This protocol describes a method for microinjection of mouse zygotes to produce transgenic mice. - [Read Microinjection of Mouse Zygotes Protocol]

Category: Microinjection Protocols: Microinjection Setup Protocols

This protocol describes the microinjection setup for injecting mouse zygotes to produce transgenic mice. - [Read Microinjection Setup Protocol]

Category: Mouse Protocols

For all the transgenic or other mouse researchers out there who have had to explain why colony infections are not just bad for the welfare of the animals concerned, but may invalidate the ongoing studies as well. - [Read Natural Pathogens of Laboratory Mice and Rats]

Category: Mouse Protocols: Mouse Embryology Protocols

Considerations in planning an effective mouse colony for transgenic production. Includes: Mouse strain; Colony size; Efficiency of superovulation; Parental Suitability; Pseudo-pregnancy. - [Read Planning an Effective Mouse Colony]

Category: Mouse Protocols: Mouse Genetics Protocols

Method is used chiefly to genotype transgenic and knockout mice. Each 6-10-mm snippet of mouse tail yields 50-100 µg of DNA that can be used in dot or slot blotting to detect a transgene of interest, in Southern hybridization to detect DNA fragments that are <20 kb in size, and as a template in PCRs. - [Read Preparation of Genomic DNA from Mouse Tails and Other Small Samples Protocol]

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: Genomic DNA Isolation Protocols

Simple protocol is used to extract DNA from small numbers of cultured cells and from fragments of soft or bony tissues.  The method is used chiefly to genotype transgenic and knockout mice.  Each 6-10-mm snippet of mouse tail yields 50-100 µg of DNA that can be used in dot or slot blotting to detect a transgene of interest, in Southern hybridization to detect DNA fragments that are <20 kb in size, and as a template in PCRs. - [Read Preparation of Genomic DNA from Mouse Tails and Other Small Samples Protocol]

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]

Category: Transgenic Mouse Protocols: Mouse Genotyping Protocols

PCR screens must be designed to detect transgene DNA at the single copy level.Copy standards are prepared by mixing non-transgenic tail DNA with a known amount of transgene DNA to produce transgene copy standards. University of Michigan Transgenic Animal - [Read reparation of Copy Standards for Southern Blot Copy Number Determination]

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

Arabidopsis can be stably transformed using Agrobacterium tumefaciens-mediated transfer of T-DNA. We describe the generation of transgenic plants via root transformation in tissue culture, which can be useful for transforming sterile mutants. - [Read Root Transformation of Arabidopsis Protocol]

Category: Mouse Protocols: Mouse Genetics Protocols

Protocol describes a simple procedure for DNA fragment isolation and purification. The resulting DNA fragment can then. be used for microinjection to produce transgenic mice - [Read Simple, Reliable Steps for DNA Fragment Isolation and Purification Protocol]

Category: Plant Biology Protocols

Protocol includes: Seed Sterilization Step; Seed Germination Step; Agrobacterium Inoculum Preparation; Explant Preparation; Co-cultivation Plates; Shoot Initiation Step; Shoot Elongation Step; Rooting Step. - [Read Soybean Cotyledonary-node Agrobacterium-mediated Transformation System]

Category: Transgenic Mouse Protocols

Timeline for Transgenic Mouse Analysis and production. University of Michigan Transgenic Animal Model Core. Transgenic Animal Web. - [Read Timeline for Transgenic Mouse Analysis]

Category: Molecular Model Organisms: Mouse Animal Protocols: Mouse Genotyping Protocols

Description:, Universal Mouse Genotyping Protocol, Primer Design, PCR Cycling, HotSHOT Preparation of Murine DNA for PCR. WUSM. Mouse Genetics Core. ... - [Read Transgenic - Universal Mouse Genotyping Protocol Using PCR]

Category: Transgenic Mouse Protocols

Mouse Breeding Suggestions, ES Cell-Mouse Chimera Breeding, Record Keeping, Transmission Patterns. University of Michigan Transgenic Animal (Web) Model Core - [Read Transgenic Mouse Breeding]

Category: Transgenic Mouse Protocols

Clone the Transgene, transgene Screening, Transgene Expression, DNA microinjection, Identify Transgenic Founders, Transgenic Analysis. Transgenic Animal Web. - [Read Transgenic Outline: Mice and Rats]