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Breaking the Tips of Embryonic Stem (ES) Cell Injection Needles Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4417

Category: Stem Cell Protocols: Stem Cell Injection Protocols

Injection needles require sharp tips that allow the blastocyst to be penetrated easily. This protocol describes a method of creating sharp injection needle tips. - [Read Breaking the Tips of Embryonic Stem (ES) Cell Injection Needles Protocol]

C. elegans Worm Injection Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=6

Category: C. elegans Protocols: C. elegans Injection Protocols

Protocol for C. elegans worm injection. - [Read C. elegans Worm Injection Protocol]

Differences in Injection of X. laevis and X. tropicalis - http://tropicalis.berkeley.edu/home/manipulate_embryos/microinjection/differences.html

Category: Xenopus Protocols: Xenopus Injection Protocols

Differences in injection of X. laevis and X. tropicalis. Includes: X. tropicalis lays eggs about 4 hours after a boost of hCG; In vitro fertilization is not nearly as efficient in trops compared to laevis; X. tropicalis embryos are much softer than X. laevis embryos; X. tropicalis embryos whose jelly coats are removed by cysteine have a loose sticky vitelline membrane; X. tropicalis do not have a "summer slump". - [Read Differences in Injection of X. laevis and X. tropicalis]

ES Cell Injection Into Mouse Embryos - http://www.oup.co.uk/pdf/pas/13-10-4.pdf

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

ES Cell Injection Into Mouse Embryos. Practical Approach Online, Oxford University Press. - [Read ES Cell Injection Into Mouse Embryos]

Gene Delivery by Direct Injection (Microinjection) Using a Controlled-Flow System Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4654

Category: Microinjection Protocols: Microinjection of DNA Protocols

This protocol describes a method for constant-flow microinjection using the Pneumatic PicoPump (World Precision Instruments). This type of system is very simple and can be assembled on a relatively low budget. In this method, a constant flow of sample is delivered from the tip of the pipette, and the amount of sample injected into the cell is determined by how long the pipette remains in the cell. - [Read Gene Delivery by Direct Injection (Microinjection) Using a Controlled-Flow System Protocol]

Gene Delivery by Direct Injection (Microinjection) Using a Pulsed-Flow System Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4653

Category: Microinjection Protocols: Microinjection of DNA Protocols

This protocol describes a method for pulsed-flow microinjection using the Eppendorf FemtoJet injector and Eppendorf InjectMan; this is the most common type of pulsed-flow microinjection system currently being used. The advantage of this type of system over a controlled-flow system is that much more control is available over the injection parameters, reducing variability in injections. In addition, the system allows a diagonal insertion of the needle into the cell. - [Read Gene Delivery by Direct Injection (Microinjection) Using a Pulsed-Flow System Protocol]

Hydrodynamics-Based Gene Transduction Protocol - http://www.natureprotocols.com/2006/06/29/hydrodynamicsbased_gene_transd.php

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]

Immunoprecipitation of Homer 1a, Injection of Virions and In Situ Hybridization in the Spinal Cord - http://www.natureprotocols.com/2006/06/30/immunoprecipitation_ip_of_home.php

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol for the immunoprecipitation (IP) of Homer 1a, injection of virions and in situ hybridization in the spinal cord. Includes: Immunoprecipitation (IP) of Homer1a from spinal cord; Injection of virions in the parenchyma of the spinal dorsal horn in vivo; Generation of cRNA probes; Analysis of DIG-dUTP incorporation; Tissue hybridization. - [Read Immunoprecipitation of Homer 1a, Injection of Virions and In Situ Hybridization in the Spinal Cord]

Injection of dsRNA and Electroporation in Avian Embryos Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4504

Category: Avian Bird Protocols

Injection of dsRNA and Electroporation in Avian Embryos Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4504

Category: RNAi Technology Protocols: RNAi Avian Embryos Protocols

Protocol describes a method for in ovo transfection of avian embryos with double-stranded RNA (dsRNA). The dsRNA is injected into the spinal cord of the embryo. Subsequent electroporation facilitates the cellular uptake of the dsRNA molecules. It may be necessary to optimize the stage of the embryo and the electroporation procedure to improve the effectiveness of in ovo RNAi—cell competence changes with differentiation. - [Read Injection of dsRNA and Electroporation in Avian Embryos Protocol]

Injection of ES Cells Into Morulae - http://www.oup.co.uk/pdf/pas/13-10-5.pdf

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

Injection of ES Cells Into Morulae. Practical Approach Online, Oxford University Press - [Read Injection of ES Cells Into Morulae]

Intraperitoneal (IP) Injection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4352

Category: Mouse Protocols: Mouse Surgical Procedures Protocols

Protocol describes intraperitoneal (IP) injection, which is the typical means of introducing most compounds, such as hormones and anesthetics, into the mouse. - [Read Intraperitoneal (IP) Injection Protocol]

Introduction of Double-Stranded RNA in C. elegans by Injection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4315

Category: C. elegans Protocols: C. elegans Genetics Protocols

Double-stranded RNA (dsRNA) can be efficiently introduced into Caenorhabditis elegans by microinjection into the gonad, the gut, or the body fluid. The RNAi effect will spread within the nematode, exerting an effect beyond the site of injection. - [Read Introduction of Double-Stranded RNA in C. elegans by Injection Protocol]

Introduction of Double-Stranded RNA in C. elegans by Injection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4315

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Introduction of Double-Stranded RNA in C. elegans by Soaking Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4316

Category: C. elegans Protocols: C. elegans Genetics Protocols

Double-stranded RNA (dsRNA) can be introduced into Caenorhabditis elegans by soaking the animals in a solution of dsRNA. Alternative methods are dsRNA injection (see Introduction of Double-stranded RNA in C. elegans by Injection) and feeding the animals with bacteria that produce dsRNA. - [Read Introduction of Double-Stranded RNA in C. elegans by Soaking Protocol]

Introduction of Double-Stranded RNA in C. elegans by Soaking Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4316

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Double-stranded RNA (dsRNA) can be introduced into Caenorhabditis elegans by soaking the animals in a solution of dsRNA. Alternative methods are dsRNA injection and feeding the animals with bacteria that produce dsRNA. - [Read Introduction of Double-Stranded RNA in C. elegans by Soaking Protocol]

Making Injection Pipettes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4395

Category: Microinjection Protocols: Microinjection Equipment

This protocol describes the determination of useful settings with the Sutter Puller P-97 to make injection pipettes for microinjection. - [Read Making Injection Pipettes Protocol]

Making Injection Pipettes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4395

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol describes the determination of useful settings with the Sutter Puller P-97 to make injection pipettes for microinjection. - [Read Making Injection Pipettes Protocol]

Microinjecting C.elegans Worms Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=5

Category: C. elegans Protocols: C. elegans Injection Protocols

Protocol for microingecting C.elegans worms. Includes: Making the DNA solution; Setting up the scope, loading the needle, mounting, and breaking the needle; Mounting worms on an injection pad; Injecting. - [Read Microinjecting C.elegans Worms Protocol]

Microinjection of Mouse ES Cells into Blastocysts - http://www.bcm.edu/dtmc/?pmid=7791

Category: Microinjection Protocols: Microinjection of DNA Protocols

The core defines one injection as a total of forty blastocysts injected on two consecutive days using one or two clones for the same mutation. Please, refer to the Pricing page for information on the cost of one injection. - [Read Microinjection of Mouse ES Cells into Blastocysts]

Plating Cells for Microinjection Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4650

Category: Cell Culture Protocols

Protocol describes a method for plating cells for microinjection onto etched coverslips. The coverslips for microinjection must be marked so that microinjected cells can be identified at time points after injection. - [Read Plating Cells for Microinjection Protocol]

Polyclonal Antibody Production Protocol - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Immunology/PAb.html

Category: Antibody Protocols: Antibody Production Protocols

Protocol for polyclonal antibody production. Very useful for rapid and simple generation of antibodies for western blots, ELISA assays, and immunoprecipitation. Includes: Rabbit Immunization; Initial Preparation; Pre-bleed; Antigen Injection; Monitoring of Titer; Purification of Antibodies. - [Read Polyclonal Antibody Production Protocol]

Preparation of Embryonic Stem (ES) Cells for Injection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4416

Category: Stem Cell Protocols: Stem Cell Injection Protocols

Injection of ES cells into the cavity of blastocysts is one of the main methods for generating chimeras. This protocol describes the preparation of ES cells for injection. - [Read Preparation of Embryonic Stem (ES) Cells for Injection Protocol]

Preparing Injection Pads Protocol - http://www.biochem.northwestern.edu/ibis/morimoto/research/Protocols/IX.%20C.%20elegans/A.%20General/6.%20injection%20pads.pdf

Category: C. elegans Protocols: C. elegans Injection Protocols

Protocol for preparing injection pads. - [Read Preparing Injection Pads Protocol]

Pristionchus Pacificus Genetic Protocols - http://www.wormbook.org/chapters/www_ppageneticprotocols/ppageneticprotocols.pdf

Category: Developmental Biology: Utilizing Model Organisms Protocols

Protocols on the genetics of Pristionchus pacificus. Includes: Freezing worms; Mutagenesis; Construction of deletion libraries to generate P. pacificus gene knock-outs; RNAi and morpholino by injection. - [Read Pristionchus Pacificus Genetic Protocols]

Articles (1-6 of 6)

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

Microinjection Pipettes Protocol

Microinjection Pipettes Preparation Protocol.

Acid Washing Microinjection Pipettes Protocol

Acid Washing of Glass microinjection pipettes protocol.

Microinjection Pipette Pulling Protocol

A simple protocol for microinjection pipette pulling.

Coupling Cysteine Containing Peptides to Carrier Protein for Immunization and Polyclonal Antibody

This protocol is used in the production of polyclonal antibodies that recognize a peptide sequence of interest.

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.

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