Navigation Links
Measuring siRNA-mediated knockdown of IL-8 mRNA

By Genospectra, Inc., 6519 Dumbarton Circle, Fremont, CA 94555.



A critical step in siRNA-mediated gene expression knockdown studies is the validation of expression inhibition. The QuantiGene Reagent System and the Molecular Devices LMax II384 microplate luminometer were used to measure IL-8 mRNA in PMA-induced HeLa cells, following siRNA treatment. Quantitative assessment of the knockdown discerned differences of less than 10% in mRNA expression levels.


INTRODUCTION
There are many sources of experimental variation that make accurate quantification of mRNA difficult when standard methods are employed. Potency and stability of siRNA coupled with variation in transfection efficiencies lead to varying effectiveness of knocking down of the target genes mRNA(1, 2) and, thus, to variability observed in RNA interference (RNAi) experiments.2 Compounding these factors are variability and inaccuracy arising from differences in sample loss during the RNA purification steps, as well as sequence-specific and random biases that occur during enzymatic amplification processes. This application note presents the QuantiGene Reagent System as an alternative method for direct RNA quantification directly from cell lysates. The siRNA-mediated knockdown of IL-8 gene expression in induced HeLa cells is employed as a model system.

QuantiGene assay principle
Bypassing many of these variability issues, the QuantiGene assay allows quantification of RNA levels directly from fresh or fixed cell lysates or tissue homogenates. This technology, based on branched DNA, allows for amplification of the signal, rather than the target, following the cooperative hybridization between the target mRNA and a gene-specific Probe Set.3 Signal amplification is precisely controlled by the Probe Set design and, therefore, is more reproducible than polymerase
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Measuring multiple apoptosis parameters with the Agilent 2100 bioanalyzer
2. Measuring Gene Silencing Effects by RT-PCR Without RNA Isolation
3. Measuring RNase Activity A Real-time Kinetic Analysis
4. Measuring Intracellular Enhanced Green Fluorescent Protein With the VersaFluor Fluorometer
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Measuring siRNA mediated knockdown mRNA

(Date:4/16/2015)... Ames, IA (PRWEB) April 16, 2015 ... (CAST) announces that the prestigious 2015 Borlaug CAST ... honored faculty member at Tuskegee University in Alabama. ... by written material; public presentations; and/or the use ... excels in all three areas. As a specialist ...
(Date:4/16/2015)... , April 16, 2015 /CNW/ - Bioenterprise Corporation is pleased ... office in Charlottetown , PEI.  Bioenterprise ... Edward Island ADAPT Council for the commercialization of agricultural ... to working with PEI ADAPT.  They have been at ... than a decade," explains Dave Smardon , President ...
(Date:4/16/2015)... April 16, 2015  Scientists from Cleave Biosciences will ... the Horizon," at the American Association for Cancer Research ... on Sunday, April 19, 2015 from 3:15pm to ... of Biology, will present new in vivo and mechanism ... inhibitor of p97, a critical enzyme that controls various ...
(Date:4/16/2015)... MENLO PARK, Calif. , April 16, 2015 /PRNewswire/ ... the Company), a biopharmaceutical company focused on developing and ... today announced that the Mayo Clinic Cancer Center ... for the ongoing, multicenter Phase I/II study of VAL-083 ... common and deadly form of human brain cancer. ...
Breaking Biology Technology:The 2015 Borlaug CAST Communication Award Goes to Channapatna Prakash 2Bioenterprise Corporation Expands into the Maritimes 2Cleave Biosciences to Participate in Special Session "New Drugs on the Horizon" at AACR Annual Meeting 2015 2Cleave Biosciences to Participate in Special Session "New Drugs on the Horizon" at AACR Annual Meeting 2015 3DelMar Pharmaceuticals Adds Fourth Clinical Trial Site in Phase I/II Multicenter Study of VAL-083 for Recurrent Malignant Glioma 2DelMar Pharmaceuticals Adds Fourth Clinical Trial Site in Phase I/II Multicenter Study of VAL-083 for Recurrent Malignant Glioma 3DelMar Pharmaceuticals Adds Fourth Clinical Trial Site in Phase I/II Multicenter Study of VAL-083 for Recurrent Malignant Glioma 4DelMar Pharmaceuticals Adds Fourth Clinical Trial Site in Phase I/II Multicenter Study of VAL-083 for Recurrent Malignant Glioma 5
... in a suit, wrinkles are almost never welcome, but two papers ... some perspective on what determines their size and shape in soft ... Amherst explore how wrinkles in a sheet adapt to an edge ... film of common plastic (polystyrene) on water and compress the sheet ...
... MADISON, Wis. , July 15 Virent Energy Systems, Inc. ... Officer and Shell executive, David Jacober , has been elected to ... , , ... Lands, End, the international apparel retailer.  She also served as CFO of ...
... , LAS VEGAS , ... , , IGEN ... assist the Board of Directors for IGEN Networks Corp. Dr Guy and Mr Celiberti,s appointments as ... , , ...
Cached Biology Technology:Virent Announces New Leadership and Board Appointments 2Virent Announces New Leadership and Board Appointments 3IGEN Networks announce the appointment of two new members to the Advisory Board 2IGEN Networks announce the appointment of two new members to the Advisory Board 3IGEN Networks announce the appointment of two new members to the Advisory Board 4
(Date:4/6/2015)... 2015 NXT-ID, Inc. (NASDAQ: NXTD ) ... focused on the growing mobile commerce market, announced ... for MINIATURE, MULTI-PURPOSE ANTENNA METHOD AND SYSTEM ... NXT-ID introduces a new revolutionary method and system ... use of miniature antenna modules for multiple purposes ...
(Date:4/2/2015)... According to preliminary estimates Fingerprint ... higher than 125 MSEK. This exceeds the guidance provided ... revenues for Q1 2015 would be in line with ... 2014. The operating result for the first quarter is ... report will as previously communicated be published on May ...
(Date:4/1/2015)... April 1, 2015   Medisafe ™, the leading ... across iOS and Android smartphones ... biofeedback into the platform to allow patients to track ... the first time, patients will be able to visualize ... impacts important biometrics, such as glucose levels and blood ...
Breaking Biology News(10 mins):NXT-ID Patents Miniature, Low Power Wireless Magnetic Stripe 2NXT-ID Patents Miniature, Low Power Wireless Magnetic Stripe 3NXT-ID Patents Miniature, Low Power Wireless Magnetic Stripe 4Fingerprint Cards Revises Revenues for Q1 2015 2Medisafe Announces Launch of Biometric Integration, Enabling Patients to Stay Adherent to Their Meds and See Tangible Health Results 2
... supporting a harmful effect of exercise on joints in the ... review of studies published in this month,s issue of the ... popular leisure-time activity in many countries throughout the Western world ... It is widely promoted in as being beneficial for weight ...
... leading provider of,embedded speech solutions today announced the ... AG, according to SVOX CEO Volker Jantzen.,SVOX has ... developed,over 25 years, including the SpeechAdvance(TM) speech recognition ... majority of the Professional,Speech Processing Group,s employees will ...
... Most people know that too much sodium from foods can ... trying to lower their blood pressure should also boost their ... Researchers found that the ratio of sodium-to-potassium in subjects, ... sodium or potassium alone. "There isn,t as much ...
Cached Biology News:SVOX Acquires Speech Processing Unit of Siemens AG, Expands Into Speech Recognition and Speech Dialog 2Cutting salt isn't the only way to reduce blood pressure 2
... be able to handle larger sample volumes, automate ... as interface with telemetry systems. The AccuSampler ... dogs, pigs, and primates. The system may ... the computer can be located outside the animal ...
The new Silencer siRNA Starter Kit is perfect for researchers new to RNAi. It provides all of the reagents, protocols, and instructions necessary to get underway with siRNA transfection and ensure su...
... A key to successful cell culture ... nutrient composition during the process, and make ... the composition. Nova BioProfile Chemistry Analyzers provide ... gases in cell culture and fermentation media. ...
... Testing Laboratory (RTL) can provide you ... years. ,,Microbial method development and validation ... the conditions of your GMP manufacturing ... products,,We offer a full spectrum of ...
Biology Products: