Navigation Links
Five Ways to Produce siRNAs

uch easier in otherwise hard to transfect cells.

Figure 3. Long Term Silencing of GFP with pSilencer 2.1-U6 hygro. HeLa cells expressing cycle 3 GFP were transfected with pSilencer 2.1-U6 hygro containing an insert encoding an siRNA targeting cycle 3 GFP or pSilencer 2.1-U6 hygro without an siRNA-encoding insert. Following transfection, the cells were selected with hygromycin. Three weeks following selection, the cells were analyzed for GFP expression by fluorescence microscopy. Green: GFP. Blue: DAPI stained nuclei. GFP levels were remarkably reduced (94%) in cells transfected with the GFP siRNA-encoding pSilencer 2.1-U6 hygro siRNA Expression Vector as compared to those transfected with an "empty" siRNA expression vector.


Method #5: siRNA Expression Cassettes
siRNA expression cassettes (SECs) are PCR-derived siRNA expression templates that can be introduced into cells directly -- without first being cloned into a vector. Initially described by Castanotto and colleagues (5), SECs include an RNA pol III promoter, a sequence encoding an siRNA hairpin, and an RNA pol III termination site. In contrast to siRNA expression vectors, which require cloning and sequencing prior to use and thus take 12 weeks to prepare, SECs can be generated by PCR in less than a day. SECs thus provide an excellent screening tool to find the most effective siRNA sequence, or to identify the most effective combination of promoter and siRNA in a given cell line. In fact, SECs provide the perfect complement to siRNA expression vectors. By incorporating restriction sites at their ends, SECs found to effectively elicit
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

Related biology technology :

1. Gram Quantities of MAb Produced with Simple Bioreactor in Serum-Free Perfusion Culture Replacing Ascitic Fluid Production
2. Silencer Validated siRNAs
3. Cenix-Designed siRNAs for 95% of Human and Mouse Genomes
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:
TAG: Five Ways Produce siRNAs

(Date:7/24/2014)... PARK, Calif. , July 24, 2014 /PRNewswire/ ... company in the emerging field of regenerative medicine, ... webcast of a presentation to investors on Tuesday, ... p.m. PDT.  The presentation will include an overview ... To access the webcast, visit http://www.ustream.tv/channel/asterias-biotherapeutics ...
(Date:7/24/2014)... of Engineering at the University of Delaware, is ... in energy engineering, environmental sustainability and electronics. , ... rock with a well-defined, crystalline structure. At the ... decided that zeolites can separate molecules with size ... (one-tenth of a nanometer), making them useful to ...
(Date:7/24/2014)... 24, 2014 SRI International has been awarded ... Institute of Allergy and Infectious Diseases (NIAID), part of ... of potential therapies for HIV infection and AIDS. The ... and AIDS and the complications and opportunistic infections associated ... sexual transmission of HIV. According to ...
(Date:7/24/2014)... July 24, 2014 WIRB-Copernicus Group ® ... and ethical review services for clinical research, announced ... a division of Richmond, VA ... biosafety and biosecurity consulting firm in ... WCG,s biosafety division – WCG Biosafety™ – will ...
Breaking Biology Technology:Asterias Biotherapeutics Announces Live Investor Webcast 2Asterias Biotherapeutics Announces Live Investor Webcast 3Asterias Biotherapeutics Announces Live Investor Webcast 4University of Delaware researcher describes new approach for creating organic zeolites 2National Institute of Allergy and Infectious Diseases Awards SRI International Contract to Study New Therapies for HIV and AIDS 2National Institute of Allergy and Infectious Diseases Awards SRI International Contract to Study New Therapies for HIV and AIDS 3WIRB-Copernicus Group Announces Acquisition of Alliance Biosciences; Executives Join WCG Biosafety Division 2WIRB-Copernicus Group Announces Acquisition of Alliance Biosciences; Executives Join WCG Biosafety Division 3
... 28, 2011 TriLink BioTechnologies, Inc. (TriLink) and Lucigen ... signed a licensing agreement for the use of TriLink,s ... MasterMix. TriLink,s CleanAmp™ dNTPs are an innovative ... standard dNTPs in a PCR assay with CleanAmp™ dNTPs, ...
... SILVER SPRING, Md., July 28, 2011 United Therapeutics Corporation ... for the quarter ended June 30, 2011. "I,m ... quarter, as well as the very good operating results we ... Executive Officer. "We remain on track to reach our 2011 ...
... Mass., July 27, 2011 SomaLogic, Inc. ... today that they have entered into an ... specific SOMAmers as critical and unique reagents ... acid amplification and polymerase chain reaction (PCR) ...
Cached Biology Technology:TriLink BioTechnologies, Inc. Announces License Agreement with Lucigen Corporation 2United Therapeutics Corporation Reports Second Quarter 2011 Financial Results 2United Therapeutics Corporation Reports Second Quarter 2011 Financial Results 3United Therapeutics Corporation Reports Second Quarter 2011 Financial Results 4United Therapeutics Corporation Reports Second Quarter 2011 Financial Results 5United Therapeutics Corporation Reports Second Quarter 2011 Financial Results 6United Therapeutics Corporation Reports Second Quarter 2011 Financial Results 7Agreement Between SomaLogic and New England Biolabs Enables the Use of SOMAmer Technology for the Optimization of Multiple PCR Products 2
(Date:7/24/2014)... specialist at The University of Texas Health Science Center ... grant to help Harris County residents whose vision problems ... Even with corrective lenses, many people with low vision ... variety of low vision devices, such as telescopes, magnifiers ... , Iyer is using the three-year, $164,645 SightFirst grant ...
(Date:7/24/2014)... week for an all-day public forum on a $1.7 ... to dredge toxic sediment from an eight-mile stretch of ... the plan, which proposes removing 4.3 million cubic yards ... one of the largest cleanups in the agency,s history. ... , "We need to ensure the future well-being of ...
(Date:7/24/2014)... To reduce fire hazard in the United States, wildland ... woody shrubs and suppressed trees (ladder fuels). These ... periods of low fire danger in order to dispose ... hazardous air pollutants, managers often cover all or part ... referred to as agricultural plastic, in order to keep ...
Breaking Biology News(10 mins):UTHealth Dr. Bhavani Iyer awarded low vision grant 2Experts Weigh the Pros and Cons of a $1.7 Billion EPA Cleanup Plan for the Passaic River at an NJIT 2Study gives new perspective on agricultural plastic, debris burning, and air quality 2
... nanotechnology fanciers suggest that, like proverbial birds of ... or clump together. This tendency, they ... manufacturing increases and the number of nanotechnology-enabled products ... Maynard, chief science advisor to the Project on ...
... completing the first large-scale report sequencing breast and colon ... studied the vast majority of protein-coding genes which now ... mutated in relatively few cancers. Their report, published ... indicates that while little is known about these less-commonly ...
... a reality today, but how can we find out about ... a full record of the Earths climate for several hundred ... that can be taken to the lab for analysis. Incredibly, ... Antarctic regions, and a European Science Foundation programme has a ...
Cached Biology News:Nanoparticle exposures happen, says expert 2Genome update defines landscape of breast and colon cancers 2Genome update defines landscape of breast and colon cancers 3European lead in reading past climates from ice cores 2European lead in reading past climates from ice cores 3
... ESGRO Complete Derivation Kit is intended ... cells from delayed blastocysts in serum-free ... Clonal Grade ... Blastocyst Incubation Medium Trypsin Solution ...
AedestaTM Animal Organ/Tissue Perfusion Soln....
RAT ANTI MOUSE JAM-2 Immunogen: Recombinant soluble JAM-2...
casein kinase 2, alpha 1 polypeptide, transcript variant 2, mRNA (cDNA clone MGC:15232 IMAGE:4121882), complete cds...
Biology Products: