Navigation Links
Confirming RNAi Effects Is Now Easier Than Ever


To validate any RNAi experiment, target mRNA reduction should be confirmed and quantitated. Validation, using a positive control siRNA that is known to be effective, is required for optimization of transfection parameters. Validation is also necessary for assessment of the effectiveness and potency of any siRNA sequence. Finally, target mRNA levels after siRNA delivery should be quantitated so that phenotypes can be correlated with the extent of mRNA knockdown.

Several different methods can be used to monitor target mRNA levels following siRNA transfection, including Northern analysis, nuclease protection assay, gel-based RT-PCR, and real-time PCR. Of these methods, real-time PCR has distinct advantages: it is the most sensitive of all of the RNA detection methods, and it can be completed in the least amount of time if optimized primer and probe sets are used.

For real-time PCR analysis of target mRNA levels for RNAi experiments, Ambion scientists recommend TaqMan Gene Expression Assays (formerly called Assays-On-Demand) from Applied Biosystems. The TaqMan Gene Expression Assays include an extensive collection of premade TaqMan primer and probe sets for quantitative, real-time PCR gene expression studies. They are considered the gold standard for gene expression analysis. Pre-designed TaqMan Gene Expression Assays are available for the human, mouse, and rat genomes, complementing Ambion's Silencer Pre-designed, Validated and Library siRNA products. Each gene specific assay is designed to work under universal thermal cycling conditions and does not require optimization prior to use.


Figure 1. Knockdown of Survivin with a Silencer Validated siRNA and Verification with TaqMan Gene Expression Assays. HeLa cells were transfected with 30 nM of a Silencer Validated siRNA targeting survivin (BIRC5) or Silencer Negative Control #1 in quadruplicate. Forty-eight hours later, two sets of samples were stained with DAPI and analyzed by fluorescence microscopy. Total RNA was isolated from the other two sample sets using RNAqueous MAG-96 and then converted to cDNA. Target cDNA levels were analyzed in duplicate using a TaqMan Gene Expression Assay for survivin (Hs00153353_m1). Data shown here represent duplicate assays from single transfections. Survivin mRNA levels were reduced 80% in the survivin siRNA treated cells relative to control siRNA treated cells. Knockdown of survivin induced morphological changes to cell nuclei. The insets show representative cells from the survivin and control siRNA treated samples.


To make it easier than ever to find TaqMan Gene Expression Assays for your siRNA experiment, Ambion's online siRNA Database now provides links to individual gene expression assays on the Applied Biosystems' website. The siRNA Database provides information about Ambion's Silencer Pre-designed siRNAs targeting >98% of all human, mouse, and rat genes in NCBI's RefSeq database, as well as Silencer Validated siRNAs, which have been functionally proven to reduce target mRNA levels by 70%. To find siRNAs and TaqMan Gene Expression Assays to your gene of interest, search our siRNA Database. You can search by gene name, RefSeq accession number, or other accession number, and will be presented with comprehensive solutions for both gene silencing and the downstream analysis of the silencing effect.

Applied Biosystems is a registered trademark of Applera Corporation. The PCR process and 5' nuclease process are covered by patents owned by Roche Molecular Systems, Inc. and F. Hoffmann-La Roche Ltd. TaqMan is a registered trademark of Roche Molecular Systems, Inc.


back to top


Ordering Information
Cat# Product Name Size 16700 Silencer Pre-designed siRNA, standard purity 20 nmol 16702 Silencer Pre-designed siRNA, standard purity 40 nmol 16704 Silencer Pre-designed siRNA, standard purity, annealed 20 nmol 16706 Silencer Pre-designed siRNA, standard purity, annealed 40 nmol 16800 Silencer Pre-designed siRNA, HPLC purified 40 nmol 16802 Silencer Pre-designed siRNA, HPLC purified 160 nmol 16804 Silencer Pre-designed siRNA, HPLC purified, annealed 40 nmol 16806 Silencer Pre-designed siRNA, HPLC purified, annealed 160 nmol 16808 Silencer Pre-designed siRNA, HPLC purified 20 nmol 16810 Silencer Pre-designed siRNA, HPLC purified, annealed 20 nmol 16900 Silencer Pre-designed siRNA, PAGE purified 25 nmol 16904 Silencer Pre-designed siRNA, PAGE purified, annealed 25 nmol
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Confirming gene silencing mechanism by pGFP/GFP22 siRNA co-transfection
2. Effects of Thermal Cycler Well-To-Well Temperature Uniformity on PCR Results
3. Measuring Gene Silencing Effects by RT-PCR Without RNA Isolation
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Confirming RNAi Effects Now Easier Than Ever

(Date:11/26/2014)... Theravalues Corporation annuncia con orgoglio il lancio di Theracurmin® ... Europe 2014 (dal 2 al 4 dicembre, ... con la maggiore biodisponibilità di sempre, con una combinazione ... norme europee. La curcumina è un pigmento ... longa ) che è stata associata con attività antiossidanti, ...
(Date:11/24/2014)... Belgium , Nov. 24, 2014 ... focused on developing diagnostic tests for cancer and other conditions, ... will be presented at the Science for Business BioWin Day ... Belgium . The data come from VolitionRx,s lung cancer ... Pneumology department of the Centre Hospitalier Universitaire (CHU) de Liege ...
(Date:11/24/2014)... (PRWEB) November 24, 2014 2014 ... Dihydrazide Industry is a professional and in-depth research ... Adipic Dihydrazide information, like its definition, classification, application, ... overview. This report covers the international market analysis, ... (such as the US, Europe, Asia, China, Japan ...
(Date:11/22/2014)... November 21, 2014 Two new educational webinars ... comes next for ALS research and how the pharmaceutical industry ... the Ice Bucket Challenge: Where Does the ALS Money Go? ... 2 December 2014, Time: 1:30pm ET, Register to ... speaker Dr. Merit Cudkowicz, Julianne Dorn Professor of Neurology at ...
Breaking Biology Technology:La curcumina con la maggiore biodisponibilità di sempre farà il suo debutto in Europa 2La curcumina con la maggiore biodisponibilità di sempre farà il suo debutto in Europa 3VolitionRx to Present Positive Data from Pilot Lung Cancer Study at BioWin Day 2014 2VolitionRx to Present Positive Data from Pilot Lung Cancer Study at BioWin Day 2014 3VolitionRx to Present Positive Data from Pilot Lung Cancer Study at BioWin Day 2014 4VolitionRx to Present Positive Data from Pilot Lung Cancer Study at BioWin Day 2014 5VolitionRx to Present Positive Data from Pilot Lung Cancer Study at BioWin Day 2014 6Adipic Dihydrazide Industry 2019 Research Forecasts on Worldwide, China Regions Now Available at ReportsnReports.com 2Adipic Dihydrazide Industry 2019 Research Forecasts on Worldwide, China Regions Now Available at ReportsnReports.com 3DrugDev Webinars in December to Explore What Comes Next for ALS Research and How We Can Make Life Easier for Clinical Sites 2
... Care Compliance Strategies, Inc. (HCCS) announced today a partnership ... to deliver a series of critical care nursing and ... Management Systems.  This training includes two separate AACN courses: ... Essentials of Nurse Manager Orientation . The courses ...
... Amsterdam Molecular Therapeutics (Euronext: AMT), a leader ... it has filed,a request for re-examination of the Marketing ... Agency.,Glybera is a gene therapy for the genetic disorder ... dossier will be completed by the end of 2011. ...
... combination of sophisticated computer modeling and advanced materials ... led by the University at Buffalo (UB) and ... (NIST), the Molecular Foundry at Lawrence Berkeley National ... simple processing flaws can seriously degrade the otherwise ...
Cached Biology Technology:HCCS Partners with American Association of Critical Care Nurses (AACN) to Offer Nurse Training 2HCCS Partners with American Association of Critical Care Nurses (AACN) to Offer Nurse Training 3Amsterdam Molecular Therapeutics Files Glybera® European Marketing Application for Re-examination 2Graphene: What can go wrong? new studies point to wrinkles, process contaminants 2
(Date:11/4/2014)... November 4, 2014   ... growth   Fuel3D , a developer of ... funding round totaling $6.4 million (£4 million). This funding builds ... earlier this year and paves the way for the commercial ... The funding round was led by Chimera Partners and ...
(Date:11/3/2014)... , Nov. 3, 2014 Research and ... "Cell Therapy - Technologies, Markets and Companies" to ... cell therapy technologies and methods, which have already started to ... stem cell transplantation is replacing the old fashioned bone marrow ... Cell therapy is bound to become a part of medical ...
(Date:11/3/2014)... and preterm birth are linked to increased risk for ... the American College of Rheumatology (ACR) journal, Arthritis ... and pre-term babies were not at greater risk of ... According to the ACR, 27 million Americans over the ... of OA range from mild to severe and include ...
Breaking Biology News(10 mins):Fuel3D Secures $6.4 Million in Expansion Funding 2Fuel3D Secures $6.4 Million in Expansion Funding 3Global Cell Therapy - Technologies, Markets and Companies: 2014 Report 2Global Cell Therapy - Technologies, Markets and Companies: 2014 Report 3Preterm, low birth-weight babies may need new hips in adulthood 2
... when it made its first U.S. appearance in New ... coast to coast, sickened more than 16,000 Americans and ... investigators hastened research to develop an effective vaccine or ... by researchers at Washington University in St. Louis points ...
... made a significant advance toward understanding the regulation of ... blood cell formation. A research study published in the ... by using adult animals to examine the role of ... cell formation. Information gained from this research will be ...
... cloning and sequencing technique has identified a surprisingly high number ... the need for closer scrutiny of public hot pools, says ... Louis and involving the University of Colorado at Boulder and ... population in a warm hospital therapy pool through a ribosomal ...
Cached Biology News:Promising new West Nile therapy cures disease in mice 2New insight into regulation of blood stem cells 2New lab technique identifies high levels of pathogens in therapy pool 2New lab technique identifies high levels of pathogens in therapy pool 3
Integrins Sampler Kit 10g each...
PKC Sampler Kit 10 mu g each...
Golgi Sampler Kit 10g each...
Umbilical cord blood eosinophils, For immunohistochemistry (IHC)...
Biology Products: