Navigation Links
High Throughput siRNA Delivery In Vitro: From Cell Lines to Primary Cells

48 hours later, GAPDH mRNA levels were monitored by real-time RT-PCR (Figure 1A). For comparison, GAPDH siRNA was delivered by a standard transfection protocol (pre-plated) in two of the cell types (Figure 1B).

Figure 1. Efficiency of Reverse Transfection in Multiple Cell Types. (A) Indicated cell lines were plated in 96 well format (8000 cells/well) and simultaneously transfected by adding transfection complexes, prepared in Opti-mem serum-free medium (Life Technologies) by mixing 0.3 l siPORT NeoFX Transfection Reagent (Ambion) and 1 nM siRNA targeting GAPDH (Silencer GAPDH siRNA, Ambion) or non-silencing control (Silencer Negative Control #1 siRNA, Ambion). 48 hours post-transfection cells were harvested and analyzed by real-time RT-PCR for both target mRNA and 18S rRNA levels. Remaining gene expression was calculated as a percentage of target mRNA in cells transfected with siRNA targeting GAPDH and cells transfected with the non-silencing control siRNA. Data was normalized against the 18S rRNA signal. Transfections were performed in duplicate. Data are presented as means SD. (B) HepG2 and HeLa cells were both reverse transfected during plating and transfected after pre-plating the cells with the siRNA targeting GAPDH or negative control siRNA. 48 hours post-transfection, GAPDH expression was measured by real-time RT-PCR. Percent gene expression was calculated as GAPDH gene expression in GAPDH siRNA transfected cells compared to those transfected with the negative control siRNA.


As shown in Figure 1, the reverse transfection method provided significant reduction in GAPDH
'"/>

Source:


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

Related biology technology :

1. Innovative Tissue Array Technology for High-Throughput Screening of Gene Expression
2. Highest Possible Transformation Efficiencies for High-Throughput Applications
3. High-Throughput Cultivation of Bacterial Clones
4. High-Throughput Isolation of Total RNA
5. High-Throughput System Generates Ultra-Pure PCR Products Suitable for All Applications
6. The DIG System Nonradioactive Automated High-Throughput In Situ Hybridization: a Powerful Tool for Functional Genomics Research
7. High Throughput aRNA Amplification
8. High Throughput siRNA Electroporation
9. High Throughput RNA Isolation for High Performance Microarray Analysis
10. RNAi for High Throughput Analysis
11. Robust High Throughput RNA Isolation from Blood Samples
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: High Throughput siRNA Delivery Vitro From Cell Lines Primary Cells

(Date:9/30/2014)... Sept. 30, 2014 BioHealth Innovation, Inc. ... market-relevant biohealth innovations and increasing access to early-stage funding ... that venture capitalist, Tania Fernandez , Ph.D., has ... Fernandez will be a member of the management team ... up to $50 million in seed and early-stage equity ...
(Date:9/30/2014)... iLab Solutions announced the results of its ... on core facility operations, growth and utilization as well ... In its 4th year running, a number of long-term ... decrease in the percentage of revenue that comes from ... constraints of this trend. When asked, “What are your ...
(Date:9/30/2014)... The custom stainless steel fabrication company ... 2014. The Boston Area Chapter of ISPE (International Society ... event for Wed., Oct. 1 at Gillette Stadium Clubhouse ... largest in ISPE Boston’s long history, with more than ... expected. , HOLLOWAY AMERICA President David Simpson is ...
(Date:9/30/2014)... SoundConnect, an award winning ... to supporting business communication solutions, including business voice ... carrier services specialist has joined the ... and productive at all times. This growth is ... to cloud communications services. On a daily ...
Breaking Biology Technology:Seasoned Life Sciences Venture Capitalist Tania Fernandez Joins BHI to Help Manage New BioHealth Gap Fund 2iLab Solutions Announces the Results of the 2014 Core Facility Benchmarking Study 2iLab Solutions Announces the Results of the 2014 Core Facility Benchmarking Study 3HOLLOWAY AMERICA to Exhibit at ISPE Boston Product Show 2014 2HOLLOWAY AMERICA to Exhibit at ISPE Boston Product Show 2014 3SoundConnect Expands Carrier Services Division 2
... InterMune, Inc. (Nasdaq: ITMN ) today announced that ... October 28, 2010 at 4:00 p.m. Eastern time.  A live ... 4:30 p.m. Eastern time that same day. To ... conference ID# 20132776.  To access the webcast, please log on ...
... Oct. 21 Paragon Bioservices, a Baltimore-based CMO focused ... biologics, announced today that it has been awarded a ... Research Institute for Infectious Diseases (USAMRIID) in its vaccine ... development and scale-up production of VLP-type vaccines against Ebola ...
... The UK,s National Physical Laboratory (NPL) is working to provide ... in nanotechnology which could lead to much more accurate ... the Nobel Prize in 1986. It uses a nano-sized probe ... a finger reading Braille on an extremely small scale. The ...
Cached Biology Technology:Paragon Bioservices Awarded $4.99 Million U.S. Army Contract to Assist with Ebola Vaccine Development 2Measuring the electrical properties of nano-crystals 2
(Date:9/30/2014)... who suffer from asthma may think there,s not a ... properly taking medications and avoiding allergic triggers. , ... of Allergy, Asthma and Immunology , the scientific publication ... (ACAAI), asthma sufferers can learn lessons about managing their ... the Annals article improved her asthma once she and ...
(Date:9/29/2014)... WASHINGTON, DC September 29, 2014 -- Plants ... ,commensal, bacteria help the pants extract nutrients and ... preventing pathogens from contaminating fruits and vegetables. Now, ... commensal bacteria inside of seeds; thus ensuring that ... researchers, from the University of Notre Dame, presented ...
(Date:9/29/2014)... home should use ventilation when cooking with a gas ... a new study showed an association between gas kitchen ... , "In homes where a gas stove was used ... higher than in homes where a gas stove was ... in the College of Public Health and Human Sciences ...
Breaking Biology News(10 mins):Asthma symptoms kicking up? Check your exposure to air pollution 2Plants prepackage beneficial microbes in their seeds 2Childhood asthma linked to lack of ventilation for gas stoves, OSU study shows 2Childhood asthma linked to lack of ventilation for gas stoves, OSU study shows 3
... of the worlds main wheat crops could strike south Asias ... leaving millions to starve. The fungus, called Ug99, has spread ... If so, this is extremely bad news, as Pakistan is ... the gateway to the Asian breadbasket, including the vital Punjab ...
... platforms slated ... for delivery during 2008, ... CTHH), ( http://www.catcherinc.com ), a leading,provider of standards-based solutions that ... environments,today announced the Company has received an initial contract to ...
... - The pending federal decision about whether to protect the ... climate science as it is about climate change. The ... proposal to list the polar bear as a threatened species ... anticipated habitat loss in a warming Arctic. Climate models ...
Cached Biology News:Killer fungus spells disaster for wheat 2Killer fungus spells disaster for wheat 3Catcher Holdings Inc. Lands Contract for Initial Deployments at Nine Homeland Security and State Government Agencies 2Catcher Holdings Inc. Lands Contract for Initial Deployments at Nine Homeland Security and State Government Agencies 3Catcher Holdings Inc. Lands Contract for Initial Deployments at Nine Homeland Security and State Government Agencies 4Arctic climate models playing key role in polar bear decision 2Arctic climate models playing key role in polar bear decision 3