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Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomyc - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4399

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the preparation of feeder cell layers from MEF cells or from the STO mouse fibroblast cell line. The cells are rendered mitotically inactive by treatment with mitomycin C. The feeder layers can then be used to maintain embryonic stem (ES) cells in the undifferentiated state. - [Read Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomyc]

Preparing Mouse Embryo Fibroblasts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4398

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the preparation of mouse embryo fibroblasts (MEFs), which can then be used as feeder cells to maintain embryonic stem (ES) cells in the undifferentiated state. - [Read Preparing Mouse Embryo Fibroblasts Protocol]

Production of Mouse T Cell Hybridomas Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E663496BA646F5C4FB54D6064E1A19B&objectid=6674C004CF35A9AC863EAD85BD9F080A

Category: Immunology: T Cell Protocols

T cell hybridomas can be obtained by fusing activated T cells with tumor cells. A heterogeneous population of hybridomas can be cloned by limiting dilution to obtain hybridomas that express specificity to one T cell receptor (TCR). Protocol describes cell fusion and selection of T cell hybridomas. A protocol is provided for screening of T cell hybridomas for expression of the CD3-TCR complex by flow cytometry analysis. - [Read Production of Mouse T Cell Hybridomas Protocol]

Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4484

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the culture of embryonic stem (ES) cells using mitotically inactivating primary mouse embryonic fibroblast (MEF) cells as a feeder layer (preparation described in Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates). The ES culture medium is supplemented with recombinant leukemia inhibitory factor (LIF) to help maintain the cells as pluripotent stem cells. This protocol has been optimized for the ES-D3 cell line. - [Read Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol]

Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4484

Category: Stem Cell Protocols: Mouse Embryonic Fibroblasts Protocols

Protocol Electroporation - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4375

Category: Transfection Protocols: Electroporation Sonoporation Protocols

Protocol describes a method for introducing gene constructsinto mouse embryos by electroporation. Gene constructs can bequickly tested for tissue-specific transcriptional activityor can be used to overexpress gene products. - [Read Protocol Electroporation]

Protocols for the Maintenance of Human Embryonic Stem Cells in Feeder Free Conditions - http://www.geron.com/PDF/scprotocols.pdf

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

Includes protocols: Mouse Embryonic Fibroblasts (MEF) Preparation; Harvesting MEFs; Cryopreservation of MEFs; Thawing and maintaining MEFs; Irradiating & Plating MEFs; Culture of Human ES cells with Matrigel® and Conditioned Medium; Preparation of Conditioned Medium (CM); Preparation of Matrigel® -coated plates; Passage of human ES cells on Matrigel®; Daily maintenance of feeder-free culture; Freezing Human ES Cells; Thawing Human ES cells; Formation of Embryoid Bodies; - [Read Protocols for the Maintenance of Human Embryonic Stem Cells in Feeder Free Conditions]

Quick and Humane Sacrifice of a Mouse by Cervical Dislocation Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4357

Category: Mouse Protocols: Mouse Surgical Procedures Protocols

Protocol describes the cervical dislocation method for humane sacrifice of a mouse. - [Read Quick and Humane Sacrifice of a Mouse by Cervical Dislocation Protocol]

Removal of Zona Pellucida Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4421

Category: Mouse Protocols: Assembling Chimeras Mouse Protocols

Protocol describes a simple method for removal of the zona pellucida of preimplantation mouse embryos using an acidified Tyrode’s solution. - [Read Removal of Zona Pellucida Protocol]

RNA Isolation and RT-PCR from dsRNA-Treated Mouse Oocytes and Early Embryos Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4513

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

Protocol describes the use of RT-PCR to confirm knockdown of a gene of interest in RNAi-treated mouse oocytes and early embryos. All of the solutions used during RNA isolation and reverse transcription must be Rnase-free - [Read RNA Isolation and RT-PCR from dsRNA-Treated Mouse Oocytes and Early Embryos Protocol]

Sectioning Mouse Embryos Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4703

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol provides methods and tips for sectioning mouse embryos and transferring the sections to a microscope slide. - [Read Sectioning Mouse Embryos Protocol]

si-RNA Transfection of Mouse Bone Marrow Progenitors by Electroporation Protocol - http://www.bcm.edu/labs/goodell/protocols/siRNA_trans.pdf

Category: RNAi Technology Protocols

Protocol for si-RNA transfection of mouse bone marrow progenitors by electroporation. - [Read si-RNA Transfection of Mouse Bone Marrow Progenitors by Electroporation Protocol]

Sorting Transfected Cells Based on Gene Expression, Followed by Culture in Vivo - http://www.bdbiosciences.com/pdfs/whitePapers/23-2787-02.pdf

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Sorting Protocols

Describes how FACSort can be used to enrich for transfected mouse cells expressing high levels of the human thrombin receptor. The sorted fraction can then be cultured in vivo and reanalyzed 12 days later to show that it remains enriched for thrombin receptor-expressing cells. - [Read Sorting Transfected Cells Based on Gene Expression, Followed by Culture in Vivo]

Spectral Karyotyping Mouse Protocol - http://www.jax.org/staff/mills/protocols/spectral_karyotyping_mouse.pdf

Category: Cytogenetics Protocols: Spectral Karyotyping SKY Protocols

Protocol for spectral karyotyping mouse. - [Read Spectral Karyotyping Mouse Protocol]

Static Culture of Postimplantation Embryos for Imaging Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4459

Category: Primary Cell Isolation and Culture Protocols

This protocol describes a method for static culture of early postimplantation mouse embryos on a microscope stage. Embryos between 6.5 and 9.5 days post coitum (dpc) can be cultured and imaged for 24 hours, with very little growth retardation. - [Read Static Culture of Postimplantation Embryos for Imaging Protocol]

Static Culture of Postimplantation Embryos for Imaging Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4459

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol describes a method for static culture of early postimplantation mouse embryos on a microscope stage. Embryos between 6.5 and 9.5 days post coitum (dpc) can be cultured and imaged for 24 hours, with very little growth retardation. - [Read Static Culture of Postimplantation Embryos for Imaging Protocol]

Tail DNA for PCR (No Organic Solvents) - http://www.jax.org/imr/tail_nonorg.html

Category: Transgenic Mouse Protocols: Mouse Tail DNA Extraction Genotyping

Use mouse tail DNA directly for PCR after Proteinase K digestion. The Jackson Lab. - [Read Tail DNA for PCR (No Organic Solvents)]

The Virtual Mouse Necropsy - http://www.geocities.com/virtualbiology/

Category: Animal Research Techniques

Perform a Mouse Necropsy on the Web. Dr. J.M.Ward - [Read The Virtual Mouse Necropsy]

Timeline for Transgenic Mouse Analysis - http://www.med.umich.edu/tamc/tgtime.html

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]

Transfer of Injected Blastocysts to Pseudo Pregnant Mice Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1118

Category: Mouse Protocols: Mouse Embryology Protocols

Blastocyst transfer is usually performed 24 hours after aggregation, when the morulae have become expanded blastocysts, and on the same day as injection. A little time is given between injection and transfer to allow the blastocysts to re-expand. Includes: The Mouse Recipient; The Transfer. - [Read Transfer of Injected Blastocysts to Pseudo Pregnant Mice Protocol]

Transgenic - Universal Mouse Genotyping Protocol Using PCR - http://www.knockoutmousestation.com/mouse/detail/link-28.html

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]

Transgenic Mouse Breeding - http://www.med.umich.edu/tamc/breed.html

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]

Treat MOUSE EMBRYONIC FIBROBLASTS (MEFs) with Mitomycin C - http://www.uphs.upenn.edu/abramson/simonPlasmids.html

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

Treat MOUSE EMBRYONIC FIBROBLASTS (MEFs) with Mitomycin C - [Read Treat MOUSE EMBRYONIC FIBROBLASTS (MEFs) with Mitomycin C]

Triple Color FISH Technique for Mouse Chromosome Identification Protocol - http://info.med.yale.edu/genetics/ward/tavi/mamgenome.html

Category: Mouse Protocols: Mouse Genetics Protocols

Protocol for triple color FISH technique for mouse chromosome identification. - [Read Triple Color FISH Technique for Mouse Chromosome Identification Protocol]

Universal Mouse Genotyping Protocol - http://ddrcc.wustl.edu/Genotyping_Protocol.html

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

Universal Mouse Genotyping Protocol (downloadable pdf click here). This protocol is designed to detect sequences in the murine genome by polymerase chain ... - [Read Universal Mouse Genotyping Protocol]

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Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

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