Navigation Links
Test More siRNAs for Less


No longer must you endure the high cost and long turn-around times required for chemically synthesized siRNA. The new Silencer siRNA Construction Kit (patent pending) saves you both time and money so now you can expand your capacity for large siRNA screening experiments. More genes or more regions of the same gene can be screened for a fraction of the cost of chemically synthesized siRNA.

RNA interference (RNAi) is a powerful technique for down regulating the expression of specific genes in live cells. In mammalian cells, RNAi can be induced by transfecting small interfering RNAs (siRNAs), comprising double-stranded RNA molecules ~21 nt in length with 2 nt 3' overhangs. In order to find an siRNA with a strong effect, 35 siRNAs per gene are typically designed. This means synthesizing and purifying 610 RNA oligonucleotides. The Silencer siRNA Construction Kit produces transfection-ready siRNA at a fraction of the cost of chemical synthesis. The cost and time savings gained using the Silencer siRNA Construction Kit enable screening of more genes and more potential targets within your gene to find the most potent siRNA. Higher potency siRNA translates to better signal-to-noise ratios in your experiments and fewer nonspecific effects.


How the Kit Works
Construction of an siRNA begins with the acquisition of two inexpensive, desalted DNA oligonucleotides. As illustrated in Figure 1, these oligonucletides include an 8 base sequence complementary to the 3' end of the T7 promoter primer include d in the Silencer siRNA Construction Kit. Each gene specific oligonucleotide is annealed to the supplied T7 promoter primer. A fill-in reaction with Klenow fragment generates a double-stranded template ready for use in an in vitro transcription reaction. The two transcription reaction products are hybridized to each other, treated with DNase (to remove template) and RNase (to polish the ends of the double-stranded RNA), and column purified. The entire procedure can be completed in less than 24 hours and is easily scalable -- 15 siRNA molecules can be synthesized as easily as a single siRNA. Each transcription/purification produces enough siRNA for hundreds of transfections. The time required for double-stranded RNA synthesis and purification using the Silencer siRNA Construction Kit is a fraction of the 12 week processing time typically required for RNA oligonucleotide synthesis via phosphoramidite chemistry and subsequent purification.

siRNAs prepared using the Silencer siRNA Construction Kit were used to silence GFP expression in HeLa cells (Figure 2).

Figure 1. The Silencer siRNA Construction Kit Procedure .

Figure 2. siRNAs Prepared with the Silencer siRNA Construction Kit Efficiently Induce RNAi. Four siRNAs to different target sequences of GFP were synthesized and purified with the Silencer siRNA Construction Kit. The siRNAs were co-transfected with a plasmid expressing GFP into HeLa cells. Cells were analyzed 24 hours after tra nsfection by fluorescence microscopy. (A) GFP expression plasmid only. (B) GFP expression plasmid co-transfected with siRNA #2. (C) Silencing efficiency achieved with the 4 different siRNAs. Note the variation of siRNA potencies with different target sites.


Contents
The kit includes all necessary reagents to generate 15 purified double-stranded siRNAs ready for transfection. Also included is a detailed Instruction Manual and GAPDH control templates. The control templates can be used to verify that the kit is functional, and the GAPDH-specific siRNA made in the control reaction can be used for optimizing transfection protocols.


back to top


Ordering Information
For prices and availability, please contact our Customer Service Department.
Cat# Product Name Size 1620 Silencer siRNA Construction Kit 15 siRNA synthesis rxns
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Silencer Validated siRNAs
2. Cenix-Designed siRNAs for 95% of Human and Mouse Genomes
3. Five Ways to Produce siRNAs
4. Efficient Delivery of siRNAs to Human Primary Cells: Electroporation vs. Chemical Transfection
5. IMPROVE SILENCING SUCCESS: siRNAs THAT WORK BETTER!
6. Optimizing Chemical Transfection and Electroporation of siRNAs
7. Delivering siRNAs to Difficult Cell Types
8. Matched siRNAs and Assays: Ambion + Applied Biosystems = RNAi Success
9. Deliver siRNAs Into Primary Cells
10. Optimize Transfection of siRNAs for RNAi
11. RNAi as a Tool for Mammalian Gene Analysis: Applications of siRNAs
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:10/9/2017)... ... October 09, 2017 , ... The Giving Tree ... products targeting the needs of consumers who are incorporating medical marijuana into their ... Phoenix, Arizona. , As operators of two successful Valley dispensaries, The Giving Tree’s ...
(Date:10/7/2017)... ... October 06, 2017 , ... ... and applications consulting for microscopy and surface analysis, Nanoscience Instruments is now ... Analytical offers a broad range of contract analysis services for advanced applications. ...
(Date:10/7/2017)... , Oct. 6, 2017  The 2017 ... of three scientists, Jacques Dubochet, Joachim Frank ... developments in cryo-electron microscopy (cryo-EM) have ... within the structural biology community. The winners worked ... can now routinely produce highly resolved, three-dimensional images ...
(Date:10/6/2017)... ... October 06, 2017 , ... On Tuesday, October 24th, ABC² ... the first-ever adaptive clinical trial for glioblastoma (GBM). The featured speaker will be ... and open to the public, but registration is required. , WHAT: ABC² ...
Breaking Biology Technology:
(Date:3/29/2017)... March 29, 2017  higi, the health IT company ... North America , today announced a Series ... acquisition of EveryMove. The new investment and acquisition accelerates ... tools to transform population health activities through the collection ... higi collects and secures data today on ...
(Date:3/24/2017)... 24, 2017 The Controller General of Immigration from ... Abdulla Algeen have received the prestigious international IAIR Award for the ... Continue Reading ... ... Controller Abdulla Algeen (small picture on the right) have received the IAIR ...
(Date:3/23/2017)... DUBLIN , Mar. 23, 2017 Research ... Anti-Theft System Market Analysis & Trends - Industry Forecast to 2025" ... ... to grow at a CAGR of around 8.8% over the next ... This industry report analyzes the market estimates and forecasts for all ...
Breaking Biology News(10 mins):