mice Protocols

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]

Category: Animal Research Techniques

Anesthesia Analgesia of Mice, Rats, Rabbits, Guinea Pigs, Dogs, Cats, Macaques, Suggestions for monitoring anesthetized veterinary patients, Alternatives to the use of Methoxyflurane in Rodents, how to mix common anesthetic mixtures. Dr.Parker Univ.Iowa - [Read Anesthesia and Analgesia of Laboratory Animalsesthesia and Analgesia of Laboratory Animals]

Category: Mouse Protocols: Mouse Embryology Protocols

Information and protocol for breeding mice. - [Read Breeding Mice Protocol]

Category: Mouse Protocols

Protocol for calcium dependent by pentraxins to ligands immobilized on agarose, and effect of C-reactive protein on leptin action in mice in vivo. Includes: Testing for binding of CRP to immobilized leptin; Testing for binding of radiolabelled leptin by immobilized CRP, SAP and leptin receptor; Testing for effect of human CRP on human leptin action in mice. - [Read Calcium Dependent by Pentraxins to Ligands, and Effect of C-reactive Protein on Leptin Action]

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol describes a method for collecting blastocysts from pregnant female mice at 3.5 to 4.5 days post coitum (dpc). The blastocysts can then be injected with embryonic stem cells to make chimeras. - [Read Collecting Blastocysts Protocol]

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol describes a method for collecting zygotes from pregnant female mice. The zygotes can then be used for microinjection. - [Read Collecting Zygotes and Removing Cumulus Cells With Hyaluronidase Protocol]

Category: RNAi Technology Protocols: RNAi Mouse Embryos and Oocytes Protocols

Protocol describes a method to collect early embryos from 6-week-old mice.  Subsequently, the isolated embryos can be injected with double-stranded RNA to induce knockdown of a gene of interest. - [Read Collection of Early Mouse Embryos for RNAi Protocol]

Category: RNAi Technology Protocols: RNAi Mouse Embryos and Oocytes Protocols

Protocol describes a method to collect oocytes from 6-week-old mice.  Subsequently, the isolated oocytes can be injected with double-stranded RNA to induce knockdown of a gene of interest. - [Read Collection of Mouse Oocytes for RNAi Protocol]

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols: Rat Neuronal Cell Culture Protocols

Conditional ablation of stat3/socs3 discloses the dual role for reactive astrocytes after spinal cord injury. The current protocol demonstrates that reactive astrocytes play a crucial role in wound healing and functional recovery by using mice with a selective deletion of the signal transducer and activator of transcription-3 (STAT3) or suppression of cytokine signaling-3 (SOCS3) under the control of Nestin gene promoter/enhancer (STAT3N–/–, SOCS3N–/–). - [Read Conditional Ablation of Stat3 Socs3 Discloses the Dual Role for Reactive Astrocytes]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: MEF Protocols Mouse Embryonic Fibroblasts

WiCell Research Institute. Recipes, Frequently Asked Questions, Mouse Embryo Extraction and Tissue, Harvesting Mouse Embryonic Fibroblasts (MEF)from mice. - [Read Derivation of Mouse Embryonic Fibroblasts (MEF) Part I PDF]

Category: Mouse Protocols: Mouse Embryology Protocols

Information on the detection of estrus in mice. - [Read Detection of Estrus in Mice]

Category: Mouse Protocols

Mice fed with the cytohesin inhibitor SecinH3 for two days develop hepatic insulin resistance that can be identified by reduced liver glycogen levels, increased serum insulin and ketone body levels and decreased serum non-esterified fatty acid. To confirm the presence and identity of SecinH3 in mouse liver, we extracted the compound from liver homogenates with chloroform and identified it by LC/MS. - [Read Extraction of the SecinH3 from Mouse Liver Protocol]

Category: Real Time PCR

Genotyping Ts65Dn mice is based on doing simultaneous quantitative PCR amplification of a gene or genes in the Ts65Dn chromosome and a control gene on another chromosome (in this case Apob) and comparing the average change (delta) in threshold cycle (CT) - [Read Genotyping Mice Using Real Time]

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: Gene Delivery Protocols

This highly efficient in vivo gene transduction technique for laboratory mice. Hepatocytes are most effectively transduced by tail vein injection of a large volume of DNA solution in a short time. Practice with the injection technique is necessary!!! - [Read Hydrodynamics-Based Gene Transduction Protocol]

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: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Liver Toxicity Assays Protocols

The results of cytotoxicity tests in primary cultures of rat hepatocytes and in MDBK and McCoy cells can be used to predict the in vivo 4-wk maximum tolerated dose in rats and dogs.  A correlation between in vitro cytotoxicity, as measured in this system, and LD50 values in rats and mice has also been established. - [Read In Vitro Prediction of the Maximum Tolerated Dose Protocol]

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]

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]

Category: Mouse Protocols: Mouse Embryology Protocols

Isolation of extraembryonic tissues allows one to study normal mouse development as well as the molecular basis of defects which cause fetal death. This protocol describes a method for isolating extraembryonic membranes from pregnant mice. - [Read Isolating Mice Extraembryonic Membranes 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: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Mammalian toxicology for acute toxicity studies.  Includes: M101 Acute Limit Test (Rodents), M102 Acute Toxicity Study (Rats or Mice), M120 Dose Escalation Study (Dog), M103 Acute Dermal Limit Test, M104 Acute Dermal (LD50), Study M105 Primary Dermal Irritation, M106 Primary Eye Irritation Study, M106-A, M110 Local Lymph Node Assay in Mouse, M115 Rabbit Vaginal Irritation Assay. - [Read Mammalian Toxicology Acute Toxicity Studies]

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: Immunology

A method for ameliorating autoimmune disease by passive transfer of IVIg-primed leukocytes. Developed a method for ‘priming’ specific leukocyte populations with IVIg to study their potential role in ameliorating ITP upon passive transfer, which we describe herein. Using this technique, found that splenic CD11c+ dendritic cells reacted with IVIg, or several IVIg mimetic regimes, were primed to ameliorate ITP upon transfer to thrombocytopenic mice. - [Read Method for Ameliorating Autoimmune Disease by Passive Transfer of IVIg-Primed Leukocytes]

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]