targeting Protocols

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Electroporating DNA into Embryonic Stem (ES) Cells and Selection Methods Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4409

Category: Mouse Protocols: Mouse Genetics Protocols

Protocol describes a method for electroporating DNA into ES cells, as well as selection methods. Pilot studies should be performed to optimize the conditions for each DNA construct. The selection method described here is one of the most complex. It involves targeting constructs in which the bacterial neomycin-resistance gene disrupts the coding sequence of the mouse gene. - [Read Electroporating DNA into Embryonic Stem (ES) Cells and Selection Methods Protocol]

Embryonic Stem Cell growth media Requirements - Taconic Transgenics - http://www.taconic.com/emerging/ESCells/Growth_Media_solutions.htm

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: ES Cell Growth Media and Requirements

Reference: Michael P. Matise, Wotjek Auerbach and Alexandra L. Joyner (2000). Gene targeting: a practical approach. Protocol excerpted from Chapter 3, Production of targeted embryonic stem cell clones. Alexandra L. Joyner (ed.), 2nd edition, Oxford Unive - [Read Embryonic Stem Cell growth media Requirements - Taconic Transgenics]

ES / MEF cell culture and electroporation of targeting construct - http://ourworld.compuserve.com/homepages/TheBroons/tnp5.htm

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

ES / MEF cell culture and electroporation of targeting construct. Gary Brown - [Read ES / MEF cell culture and electroporation of targeting construct]

ES / MEF cell culture and electroporation of targeting construct - http://ourworld.compuserve.com/homepages/TheBroons/tnp5.htm

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

Dr. Gary Brown. Electroporation and transfection of ES / MEF cells using electroporation for a targeting construct. - [Read ES / MEF cell culture and electroporation of targeting construct]

ES / MEF Cell Culture and Electroporation of Targeting Construct Protocol - http://members.cox.net/microinjectionworkshop/technical/tnp5.html

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Protocol for ES/MEF cell culture and electroporation of targeting construct. - [Read ES / MEF Cell Culture and Electroporation of Targeting Construct Protocol]

ES / MEF Cell Culture and Electroporation of Targeting Construct Protocol - http://members.cox.net/microinjectionworkshop/technical/tnp5.html

Category: Stem Cell Protocols: Mouse Embryonic Fibroblasts Protocols

Protocol for ES/MEF cell culture and electroporation of targeting construct. - [Read ES / MEF Cell Culture and Electroporation of Targeting Construct Protocol]

ES / MEF Cell Culture and Electroporation of Targeting Construct Protocol - http://members.cox.net/microinjectionworkshop/technical/tnp5.html

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

Protocol for ES / MEF cell culture and electroporation of targeting construct. - [Read ES / MEF Cell Culture and Electroporation of Targeting Construct Protocol]

ES / MEF Cell Culture and Electroporation of Targeting Construct Protocol - http://members.cox.net/microinjectionworkshop/technical/tnp5.html

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol for ES / MEF cell culture and electroporation of targeting construct. - [Read ES / MEF Cell Culture and Electroporation of Targeting Construct Protocol]

Fractionation of Vacuolar and Subvacuolar vesicles and Vacuole and Cytoplasm-to-Vacuole Targeting - http://www.axis-shield.com/densityhome/optiprep/S29.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

Fractionation of (a) vacuolar and subvacuolar vesicles and (b) vacuole and cytoplasm-to-vacuole targeting (Cvt) vesicles from yeast spheroplasts in a pre-formed discontinuous iodixanol gradients. Protocol includes: Formation of yeast spheroplasts; Isolation and vesiculation of the vacuoles; Separation of the vacuolar and subvacuolar vesicles; Separation of vacuoles and Cvt vesicles from a yeast spheroplast lysate. - [Read Fractionation of Vacuolar and Subvacuolar vesicles and Vacuole and Cytoplasm-to-Vacuole Targeting]

Immunoprecipitation of mRNA-Protein Complexes Protocol - http://www.nature.com/nprot/journal/v1/n2/pdf/nprot.2006.82.pdf

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation of mRNA-protein complexes. In this protocol, an antibody targeting an RBP of interest is used to immunoprecipitate the RBP and any interacting molecules from a cell lysate. Reverse transcription followed by PCR is then used to identify individual mRNAs isolated with the RBP. This method focuses on examining an association between a specific RBP-mRNA complex, and it is best suited for a small scale screening of known or putative binding partners. - [Read Immunoprecipitation of mRNA-Protein Complexes Protocol]

PEI Nanoparticles for Targeted Gene Delivery Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/1/pdb.prot4451

Category: Gene Delivery Protocols: DNA Nanoparticles Protocols

This protocol describes the preparation of polyethylenimine (PEI)/DNA nanoparticles for targeted gene delivery. This delivery strategy improves the efficiency of gene transfer by enhancing the entry of gene vectors into the desired cells and reducing uptake by nontarget cells. We describe here methods for the conjugation of targeting peptides to PEIs, formation of DNA complexes using the conjugated PEIs or nonconjugated PEIs together with targeting peptides, and cell transfection. - [Read PEI Nanoparticles for Targeted Gene Delivery Protocol]

Precision Engineering of Plant Gene Loci by Homologous Recombination Cloning in Escherichia Coli - http://www.plantmethods.com/content/1/1/6

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol for precision engineering of plant gene loci by homologous recombination cloning in Escherichia coli. Includes: Key steps in the EL250 RED-HR locus rescue and engineering procedure; Primer design and plasmid constructs; AtSTM gap-repair construct; Targeting construct backbone; Preparation of electrocompetent EL250 cells; Transformation of BAC F24o1 and induction of recombinogenic function in EL250; AtSTM locus rescue from BAC F24o1 by gap-repair HR. - [Read Precision Engineering of Plant Gene Loci by Homologous Recombination Cloning in Escherichia Coli]

Selection of siRNA Sequences for Mammalian RNAi Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4339

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

To target a specific mRNA for degradation, a portion of the mRNA target sequence must be known, and a segment of the target mRNA must be chosen that will be used for targeting by the cognate siRNA duplex. Protocol describes the selection of siRNA sequences for mammalian RNAi. - [Read Selection of siRNA Sequences for Mammalian RNAi Protocol]

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.