Navigation Links
Optimize Transfection of siRNAs for RNAi

he optimized amount of cells (3 x 104 cells/well) and 3 l transfection reagent to establish the concentration resulting in a >50% reduction in mRNA expression. The most significant siRNA activity was attained using siRNA concentrations between 30 and 100 nM (Figure 5). We were able to achieve approximately 75%-90% reduction in GAPDH mRNA levels over control with no apparent cytotoxicity.

Figure 5. Determination of Optimal Amount of siRNA. COS-7 cells were plated at the optimized cell plating density (3.0 x104 cells per well) in a 24 well dish. 24 hours after plating, cells were transfected with 100, 30, 10, or 3 nM chemically syntheesized GAPDH siRNA or negative control siRNA using the optimized volume of transfection agent (3 L per well). 48 hours following transfection, the cells were harvested and analyzed by real-time RT-PCR for both GAPDH mRNA and 18S rRNA levels. Percent gene expression was calculated as a percentage of gene expression compared to the negative control siRNA.


Note that it is common to see relative variations in data when conducting tissue culture experiments. Reproducibility can be achieved by rigorously following protocols.

siPORT Amine is manufactured for Ambion by Mirus.

TaqMan is a registered trademark of Applied Biosystems.


REFERENCES

1. Byrom M, Pallotta V, Brown D, Ford L (2002) Visualizing siRNA in Mammalian cells: Fluorescence analysis of the RNAi ef
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9 10

Related biology technology :

1. Optimized Imaging of Protein Gels Stained with Coomassie Brilliant Blue Dye
2. Eagle Eye Optical Filter Optimized for Imaging Silver-Stained Gels
3. Optimized Dengue RT-PCR
4. Rapamycin Optimized in 100 Liter Fermentor Using BioCommand Control Strategys
5. Highest Transfection Efficiency of an Endotoxin-Sensitive Mammalian Cell Line
6. Optimizing Transfection Conditions for Studying Signal Transduction Pathways
7. Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells
8. Improve Lipid- or Calcium Phosphate-Mediated Transfection of Human Dermal Fibroblasts
9. Versatile Transfection Reagent Offers Low Toxicity and Consistent Performance
10. Low-Toxicity, Lipid-Mediated Transfection of Mammalian Cells
11. Mycoplasma Contamination Reduces the Effect of Lipid-Mediated Transfection of Mammalian Cells
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Optimize Transfection siRNAs for RNAi

(Date:7/10/2014)... 10, 2014  Franciscan St. Anthony Health-Crown ... for respiratory monitoring outside the operating room, ... in embracing state-of-the-art patient safety technology. ... breathing and can alert medical caregivers when ... amount of carbon dioxide the patient is ...
(Date:7/10/2014)... -- Research and Markets ( http://www.researchandmarkets.com/research/s3bqj2/photonic ) has ... Integrated Circuit (Monolithic Integration, Hybrid Integration, Module Integration) ... their offering. http://photos.prnewswire.com/prnh/20130307/600769 ... traces its roots in the latter half of ... went unexplored and unfulfilled for several decades owing ...
(Date:7/10/2014)... microscopic polymer light resonators that expand in the ... Photonics Laboratory have developed new optical sensors with ... sensors are ideal for detecting trace gas concentrations ... nature, and immunity to electromagnetic interference. , Although ... the MIT team conceived an extremely sensitive, compact ...
(Date:7/10/2014)... that power our laptops and electric vehicles could store ... with the help of a sponge-like silicon material. ... graphite traditionally used in one of the battery,s electrodes, ... storage capacity of graphite. A paper describing the material,s ... in Nature Communications . , "Silicon has long ...
Breaking Biology Technology:Franciscan St. Anthony Health-Crown Point Underscores Patient Safety Commitment Through Expanded Use of Capnography 2International Photonic Integrated Circuit (Monolithic Integration, Hybrid Integration, Module Integration) Market - Forecasts to 2019 2Swell new sensors 2Silicon sponge improves lithium-ion battery performance 2Silicon sponge improves lithium-ion battery performance 3
(Date:7/11/2014)... The Chinese University of Hong Kong, BGI and other ... to salt tolerance, with implication for improving this important ... online in Nature Communications provides an effective ... , Soybean is an important crop for the world. ... less genetic diversities than their wild counterparts. Among the ...
(Date:7/11/2014)... group members to reduce aggression and gain access to ... grooming activities shows a certain pattern across the day. ... Biology Letters . , Grooming between individuals in ... motives. To be groomed has hygienic benefits and is ... can provide access to infants, mating opportunities and high ...
(Date:7/11/2014)... Ume University, Sweden, have explored two different ways ... sequence leading to the formation of oxygen molecules ... in the scientific journal Nature Communications . ... day synthetic catalysts for water oxidation, which are ... the direct storage of solar energy in fuels ...
Breaking Biology News(10 mins):BGI reports a novel gene for salt tolerance found in wild soybean 2Baboons groom early in the day to get benefits later 2Molecular snapshots of oxygen formation in photosynthesis 2
... Lockheed Martin (NYSE: LMT ) has been ... of the Year" by Frost and Sullivan, recognizing the ... high levels of customer value. "Lockheed ... that play an important role our customer,s law enforcement ...
... Nitrogen is essential for plant growth. Despite the abundance ... must rely on a series of chemical reactions to ... forms. Plants have achieved this goal primarily by evolving ... nitrogen, plants, and the environment are the focus of ...
... Has a bone density scan placed you at risk ... bone-building medication? Not so fast! A University of Illinois study ... dietary calcium and vitamin D or taking calcium and vitamin ... be a last resort," said Karen Chapman-Novakofski, a U of ...
Cached Biology News:Lockheed Martin Named Biometrics "Company of the Year" by Frost and Sullivan 2Book looks at interrelationships among nitrogen, plants and the environment 2U of I study: Before you start bone-building meds, try dietary calcium and supplements 2U of I study: Before you start bone-building meds, try dietary calcium and supplements 3
Performance, mycoplasma, virus, and endotoxin tested...
... 10X BlueJuice Gel Loading ... easy loading and tracking ... agarose or native polyacrylamide ... features higher density sucrose ...
... is the most effective ... PCR products. TOPO TA ... TOPO vectors are supplied ... topoisomerase I. This enables ...
ready-to-use solution...
Biology Products: