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Bacteria-Mediated (Feeding) RNAi Protocol - http://www.genetics.wustl.edu/tslab/Protocols/Feeding_RNAi.htm

Category: C. elegans Protocols

Protocol for bacteria- mediated (feeding) RNAi. - [Read Bacteria-Mediated (Feeding) RNAi Protocol]

C. elegans RNAi Protocol - http://www.zoology.ubc.ca/~alorch/rnai-protocol.htm

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

Protocol for C. elegans RNAi. Includes: Transformation of competent cells; Blunt-end ligation; Preparation of competent cells; Dephosphorylation of linear plasmid DNA; Restriction Digest: EcoRV; Insert amplification from gDNA; Gel purification: QiaQuick gel purification kit; Mini-prep; Transformant Screening; Bacterial preparation and induction; Preparation of worms for RNAi feeding. - [Read C. elegans RNAi Protocol]

Collection of Early Mouse Embryos for RNAi Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4510

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]

Collection of Mouse Oocytes for RNAi Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4509

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]

Cosuppression in C. elegans Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4318

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Cosuppression is a process in Caenorhabditis elegans that closely resembles RNAi. In contrast to RNAi, however, the cosuppression effect in C. elegans does not spread throughout the animal. Cosuppression in C. elegans can be triggered by highly repetitive transgenes that contain gene constructs. - [Read Cosuppression in C. elegans Protocol]

Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4342

Category: Reporter Gene Assays: Luciferase Assay Protocols

It is possible that some cell lines have lost the ability to perform RNAi or that cells derived from certain tissues do not support RNAi. This reporter assay, for RNAi in mammalian cells, can be used to establish whether the cells under study are susceptible to RNAi. - [Read Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol]

Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4342

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

This reporter assay, for RNAi in mammalian cells, can be used to establish whether the cells under study are susceptible to RNAi. - [Read Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol]

Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4345

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

Protocol describes the use of western blotting to analyze the RNAi-induced depletion of target protein from mammalian cells. Immunofluorescence can be used as an alternative. - [Read Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol]

Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4345

Category: Western Blot Protocols

dsRNA Synthesis Protocol - http://flyrnai.org/RNAi_FAQ_dsrna_syn.html

Category: siRNA and RNAi Protocols

dsRNA From Clones, Primer Designed dsRNA. Drosophila RNAi Screening Center at Harvard Medical School - [Read dsRNA Synthesis Protocol]

DSRNAI EXPERIMENTS IN DROSOPHILA S2 CELL CULTURE - http://www.mann.embl-heidelberg.de/GroupPages/PageLink/activities/iTAP/Experimental/dsRNAi_Dixon.html

Category: siRNA and RNAi Protocols

DSRNAI EXPERIMENTS IN DROSOPHILA S2 CELL CULTURE. using T7 to create RNA. Carolyn Worby. Dixon Lab RNAi experiments. - [Read DSRNAI EXPERIMENTS IN DROSOPHILA S2 CELL CULTURE]

Electroporation and RNAi in the Rodent Retina In Vivo and In Vitro Protocol - http://genetics.med.harvard.edu/~cepko/electroporation/PNAS-final.pdf

Category: Transfection Protocols: Electroporation Sonoporation Protocols

Protocol for electroporation and RNAi in the rodent retina in vivo and in vitro. - [Read Electroporation and RNAi in the Rodent Retina In Vivo and In Vitro Protocol]

Electroporation and RNAi in the Rodent Retina In Vivo and In Vitro Protocol - http://genetics.med.harvard.edu/~cepko/electroporation/PNAS-final.pdf

Category: RNAi Technology Protocols

Protocol for electroporation and RNAi in the rodent retina in vivo and in vitro. - [Read Electroporation and RNAi in the Rodent Retina In Vivo and In Vitro Protocol]

Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4344

Category: Immunology: Immunofluorescence Protocols

This protocol describes the use of a specific antibody that recognizes the targeted gene product to detect RNAi-induced gene knockdown in mammalian cells. Western blot technology can be used as an alternative (see Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting). - [Read Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol]

Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4344

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

Protocol describes the use of a specific antibody that recognizes the targeted gene product to detect RNAi-induced gene knockdown in mammalian cells. Western blot technology can be used as an alternative. - [Read Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol]

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]

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

Lactose RNAi Plates: Minimal Lactose Media Protocol - http://www.genetics.wustl.edu/tslab/Protocols/Lactose_RNAi_Plates.htm

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol for lactose RNAi plates: minimal lactose media. - [Read Lactose RNAi Plates: Minimal Lactose Media Protocol]

Mammalian Cell Culture and Preparation of Cells for RNAi in 24-Well Plates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4341

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

This protocol introduces a method for maintaining mammalian cell culture suitable for treatment with siRNA. Transfection of cultured cells with siRNAs (see Transfection of siRNA Duplexes into Mammalian Cells) and downstream analysis of the knockdown cells. - [Read Mammalian Cell Culture and Preparation of Cells for RNAi in 24-Well Plates Protocol]

Mammalian Cell Culture and Preparation of Cells for RNAi in 24-Well Plates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4341

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

Protocol introduces a method for maintaining mammalian cell culture suitable for treatment with siRNA. Transfection of cultured cells with siRNAs and downstream analysis of the knockdown cells. - [Read Mammalian Cell Culture and Preparation of Cells for RNAi in 24-Well Plates Protocol]

Microinjection of dsRNA into Mouse Oocytes and Early Embryos Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4511

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

Protocol describes how to introduce a double-stranded RNA (dsRNA) of choice into mouse oocytes or fertilized one-cell embryos by microinjection. For collection of mouse oocytes and early embryos, see Collection of Mouse Oocytes for RNAi and Collection of Early Mouse Embryos for RNAi. - [Read Microinjection of dsRNA into Mouse Oocytes and Early Embryos Protocol]

Preparation and Fractionation of Drosophila Embryo Extracts for the In Vitro Characterization of RNA - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4324

Category: RNAi Technology Protocols: RNAi Drosophila Protocols

Protocol describes a method to prepare and fractionate Drosophila embryo extracts. These extracts can be used for the in vitro characterization of RNAi. - [Read Preparation and Fractionation of Drosophila Embryo Extracts for the In Vitro Characterization of RNA]

Preparation of dsRNA Molecules for RNAi in Mouse Oocytes and Early Embryos Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4508

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

Protocol describes in vitro transcription with the SP6 polymerase. For other polymerases (T3, T7), simply use the appropriate buffer and desired RNA polymerase instead of the SP6 polymerase. - [Read Preparation of dsRNA Molecules for RNAi in Mouse Oocytes and Early Embryos Protocol]

Preparation of siRNAs from Drosophila Embryo Extracts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4325

Category: RNAi Technology Protocols: RNAi Drosophila Protocols

siRNAs produced upon the addition of dsRNA to Drosophila embryo extract are enriched in a micrococcus-nuclease-resistant fraction. After proteinase K treatment and dephosphorylation with calf intestinal phosphatase, these siRNAs mediate efficient RNAi in vitro. - [Read Preparation of siRNAs from Drosophila Embryo Extracts Protocol]

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