sirna Protocols

Category: RNAi Technology Protocols

Protocol describes a procedure to anneal sense and antisense siRNA strands to form a duplex.  The duplexes can then be transfected into cultured cells. - [Read Annealing siRNAs to Produce siRNA Duplexes Protocol]

Category: Cloning Protocols: Vector Protocols

Protocol for cloning siRNA duplexes into pSUPER or pSUPER retro vectors. - [Read Cloning siRNA Duplexes into pSUPER or pSUPER Retro Vectors Protocol]

Category: RNAi Technology Protocols

Protocol for cloning siRNA duplexes into pSUPER or pSUPERretro vectors. - [Read Cloning siRNA Duplexes into pSUPER or pSUPERretro Vectors Protocol]

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]

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]

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Protocol uses Rnase protection to detect short interfering RNAs (siRNAs) in RNA preparations from Caenorhabditis elegans.  SiRNAs can also be detected by northern blot.  However, the Rnase protection assay seems to be more sensitive. - [Read Detection of siRNA in C. elegans Using Rnase Protection Protocol]

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]

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]

Category: RNAi Technology Protocols

Short interfering RNAs (siRNAs) can be used to prime RNA synthesis by the RNA-dependent RNA polymerase (RdRP).  SiRNAs can be used by RdRP as primers for specific cellular mRNAs, forming dsRNA products capable of inducing transitive RNAi. - [Read Protocol for siRNA-Primed RNA Synthesis Protocol]

Category: siRNA and RNAi Protocols

RNAi Interference Resources - Ambion. Introduction, product database for siRNA and RNAi. Ambion. - [Read RNAi Interference Resources - Ambion]

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]

Category: RNAi Technology Protocols

Protocol Involves the transfection of siRNA into RAW 264. 7 cells using Lipofectamine 2000.  Cells are transfected with siRNA twice (on subsequent days).  Target gene knockdown is assessed from total RNA isolated 48 hr post-transfection or from protein isolated 72 hr post-transfection. - [Read siRNA Double Transfection of RAW 264.7 Cells with Lipofectamine Protocol]

Category: RNAi Technology Protocols

Protocol Involves the transfection of siRNA into RAW 264. 7 cells using Lipofectamine 2000.  Cells are transfected with siRNA twice (on subsequent days).  Target gene knockdown is assessed from total RNA isolated 48 hr post-transfection or from protein isolated 72 hr post-transfection. - [Read siRNA Double Transfection of RAW 264.7 Cells with Lipofectamine Protocol II]

Category: siRNA and RNAi Protocols

siRNA protocols - siRNA transfection. Mellor Lab - [Read siRNA protocols - siRNA transfection]

Category: siRNA and RNAi Protocols

siRNA Transfection Protocol. - [Read siRNA Transfection Protocol]

Category: siRNA and RNAi Protocols

siRNA Transfection Protocol for TransPass R1 NEB - [Read siRNA Transfection Protocol for TransPass R1 NEB]

Category: siRNA and RNAi Protocols

Selection of siRNA duplexes from the target mRNA sequence, Preparation of siRNA duplexes, Useful siRNA reagent combinations, Transfection of siRNA duplexes, Sequences of siRNA duplexes used in our studies. Tuschl Lab. - [Read The siRNA user guide]

Category: RNAi Technology Protocols

The siRNA user guide. Includes: Selection of siRNA duplexes from the target mRNA sequence; Preparation of siRNA duplexes; Useful siRNA reagent combinations; Transfection of siRNA duplexes; Sequences of siRNA duplexes used in our studies. - [Read The siRNA User Guide]

Category: Transfection Protocols

Protocol describes a method for the delivery of siRNAs into mammalian cells in the absence of reporter plasmids. This is best achieved with transfection reagents developed for the delivery of antisense oligodeoxynucleotides. The quantities of reagents given below are calculated for the transfection of one well of a 24-well plate. - [Read Transfection of Mammalian Cells with siRNA Duplexes Protocol]

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

Protocol describes a method for the delivery of siRNAs into mammalian cells in the absence of reporter plasmids.  This is best achieved with transfection reagents developed for the delivery of antisense oligodeoxynucleotides.  The quantities of reagents given below are calculated for the transfection of one well of a 24-well plate. - [Read Transfection of Mammalian Cells with siRNA Duplexes Protocol]