Navigation Links
Detecting miRNA & siRNA

nger (29 nt) than the protected fragment generated by the mir-16 probe (22 nt). These probes were then used to simultaneously detect miR-16 miRNA and GAPDH mRNA in total RNA from HeLa cells (Figure 3). The results indicate that both targets are easily detected, and that surprisingly, the level of miR-16 in HeLa cells is greater than that of GAPDH mRNA, a transcript that is known to be highly abundant.


Simultaneous Detection of siRNA Expression and Target Gene Knockdown
For the development of improved siRNA delivery systems and siRNA expression vectors, particularly those that permit targeted delivery or tissue specific expression, researchers need to monitor siRNA levels -- preferably in conjunction with the assessment of target transcript down regulation. To determine if the solution based assay permitted correlation of expressed siRNA levels and targeted mRNA knockdown, probes were generated to a GAPDH siRNA known to effectively reduce GAPDH expression, and to the GAPDH mRNA target. An expression vector (pSilencer 2.0-U6) engineered to express either the GAPDH siRNA or a scrambled control siRNA was transfected into HeLa cells. Three days after transfection, total RNA was isolated and the solution hybridization assay was performed. Using the assay, it was possible to simultaneously detect siRNA expressed from the vector (Figures 3 and 5A), as well as the GAPDH mRNA (Figure 5B). As expected, the GAPDH mRNA levels were found to be reduced considerably (~60%) in cells transfected with the GAPDH siRN
'"/>

Source:


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

Related biology technology :

1. Detecting Gastrin-Releasing Peptide Receptor by in situ PCR on Archived Tissue
2. Tools for Detecting MSH2 Expression in Chinese Hamster Ovary Cells
3. Two-dimensional Optimization of a Semi-nested PCR for Detecting Listeria monocytogenes
4. Detecting Attomole Amounts of Small RNA
5. Cancer-Related miRNAs Uncovered by the mirVana miRNA Microarray Platform
6. miRNA Expression in White Blood Cells
7. Isolate It All: siRNA miRNA Total RNA Native Protein
8. Get the miRNA OutPlant Applications
9. Novel MicroRNA Array Technology for Sensitive miRNA Profiling
10. pSilencer 4.1-CMV: Versatile Vectors for Expression of siRNA, miRNA, and mRNA
11. Precursor miRNAs for Successful miRNA Functional Studies
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Detecting miRNA siRNA

(Date:7/24/2008)... -- The global pharma,ingredients and services com... returns this year to Frankfurt on Tuesday 30 Sept...ICSE contract services and P-MEC machinery and,equ...y located in the,adjoining Messe Halls 3, 4, 5 and...ts and 2008 marks the fifth time that the city hos...
(Date:7/24/2008)...swire-FirstCall/ -- Poniard,Pharmaceuticals, Inc. ...sed,on oncology, today announced that it will hold... quarter results and provide a corporate update,i...n for picoplatin, the,Company,s lead product candi...cutive officer, Ronald A. Martell, president and c...
(Date:7/24/2008)...s Greater Control of Common, Pests and Provides R...s, ST. LOUIS, July 24 /- T...has deregulated MON 89034, a new insect,protection...ommercially, this,new above ground insect control ...ground and Roundup Ready technologies to be sold a...
(Date:7/24/2008)..., July 24,/- Rosetta Genomi...ent of microRNA-based diagnostics and therapeutics...ed the acquisition of Parkway,Clinical Laboratorie...g a,CLIA-certified lab located in Bensalem, Pennsy...000, consisting of $1,900,000 in cash and,$1,000,0...
Breaking Biology Technology:Frankfurt Hosts Continuing Growth of CMPi Pharma Events 2Frankfurt Hosts Continuing Growth of CMPi Pharma Events 3Poniard Pharmaceuticals to Host Second Quarter 2008 Financial Results Conference Call on July 31 2New Insect Protection Technology From Monsanto Company Deregulated by USDA 2New Insect Protection Technology From Monsanto Company Deregulated by USDA 3Rosetta Genomics Completes Acquisition of Parkway Clinical Laboratories Inc. 2Rosetta Genomics Completes Acquisition of Parkway Clinical Laboratories Inc. 3Rosetta Genomics Completes Acquisition of Parkway Clinical Laboratories Inc. 4Rosetta Genomics Completes Acquisition of Parkway Clinical Laboratories Inc. 5
...Information Networks was founded in 1993, founde...ing principles: high quality customer service and ... channeled these philosophies to create one of the... providers in the Midwest. The company refuses to ... their staff and adding to their service options. ...
...chnology professionals in Wisconsin announced on M... Association of Wisconsin, a forum for informatio...erates more than $14B in revenue and hires more th...,The association has 30 founding members, of which...are a mix of law, investment, utility and financia...
...t at hiring and firing workers. Great businesses h...ent in ways that continuously grow their experienc...ements. Creating work environments that promote pe...ries is the next imperative for companies seeking ... is surprising how few companies develop and move ...
...oe Hogan, head of GE Healthcare Technologies in Wa...hcare in London next year as part of the parent c...o six. , ,Hogan will succeed Bill Castell as head ...sident and CEO of the Waukesha-based GE Healthcare...ed Jeff Immelt after Immelts promotion to succeed ...
Other Biology Technology:Berbee puts people first to deliver high-quality technology 2Berbee puts people first to deliver high-quality technology 3Berbee puts people first to deliver high-quality technology 4Statewide IT association will address IT issues and priorities 2Statewide IT association will address IT issues and priorities 3Agility is the next talent management imperative 2Agility is the next talent management imperative 3Agility is the next talent management imperative 4
(Date:7/24/2008)...e researchers have found a molecular link between ...ism. The discovery reveals new possibilities for t...d diseases. , Paolo Sassone-Corsi, Distinguished...eagues have identified that an essential protein c...ythms, works in balance with another protein calle...
(Date:7/24/2008)...08. A fire is currently burning through a study ar... about 2 years ago. Managers used data and analysi... study in the planning, analysis, and implementati...is now active., The Gotchen LSR, lies on the eas...vers about 15,000 acres of the Mount Adams Ranger ...
(Date:7/23/2008)...tions willing to collectively spend about $1 billi...half a billion metric tons of carbon dioxide per y... , It would take about that much money to put an...he world, one of the top contributors to greenhous...ted, this type of program could have potential to ...
(Date:7/23/2008)...In a study of free-living and parasitic species in...a and Baja California, a team of researchers from ...nited States Geological Survey, and Princeton Univ...se habitats exceeds that of top predators, in some...uld have significant biomedical and ecological imp...
Breaking Biology News(10 mins):Circadian rhythm-metabolism link discovered 2Landscape study may offer solutions for fire managers 2Paying to save tropical forests could be a way to reduce global carbon emissions 2Paying to save tropical forests could be a way to reduce global carbon emissions 3Paying to save tropical forests could be a way to reduce global carbon emissions 4Study shows parasites outweigh predators 2Study shows parasites outweigh predators 3BD and Direct Relief International Launch Second Volunteer Service Trip to Improve Healthcare in Ghana 4233 1BD and Direct Relief International Launch Second Volunteer Service Trip to Improve Healthcare in Ghana 4233 2BD and Direct Relief International Launch Second Volunteer Service Trip to Improve Healthcare in Ghana 4233 3Trubion Initiates Phase 1 2 Study of TRU 016 in CLL Announces Next Generation Product Candidate for RA and Provides Product Pipeline Update 1607 1Trubion Initiates Phase 1 2 Study of TRU 016 in CLL Announces Next Generation Product Candidate for RA and Provides Product Pipeline Update 1607 2Trubion Initiates Phase 1 2 Study of TRU 016 in CLL Announces Next Generation Product Candidate for RA and Provides Product Pipeline Update 1607 3Trubion Initiates Phase 1 2 Study of TRU 016 in CLL Announces Next Generation Product Candidate for RA and Provides Product Pipeline Update 1607 4Trubion Initiates Phase 1 2 Study of TRU 016 in CLL Announces Next Generation Product Candidate for RA and Provides Product Pipeline Update 1607 5FDA Probing Possible Link Between Asthma Drug and Suicide Risk 15339 1FDA Probing Possible Link Between Asthma Drug and Suicide Risk 15339 2FDA Probing Possible Link Between Asthma Drug and Suicide Risk 15339 3Insights on Paying for Boomers Care Needs in 20 40 Years Helped by New Report 15337 1Insights on Paying for Boomers Care Needs in 20 40 Years Helped by New Report 15337 2Insights on Paying for Boomers Care Needs in 20 40 Years Helped by New Report 15337 3Insights on Paying for Boomers Care Needs in 20 40 Years Helped by New Report 15337 4Insights on Paying for Boomers Care Needs in 20 40 Years Helped by New Report 15337 5
...alaria remains the subject of "best guesses," and ...l remains "driven by perceptions and politics, rat... prominent malaria researchers in PLoS Medicine. ,... Institute and University of Oxford), say that it ... of malaria endemicity was constructed, and that "...
...an be arranged along a ladder of intelligence—a so...bly at the top, followed by our close relatives, t...imy creatures. Over the past decade, this ladder h...and great social complexity in other animals. For ...hich dominant females support the rank contests of...
... fight viruses is also involved in keeping cancer ...l Center have discovered. , The anti-viral molecu...ncer cells to live, so blocking it might point to ...eport in today,s issue of Cell. , "We got the s...cell survival, even though it,s normally involved ...
...into a new life, or into new cells that may someda...in the mechanics - in how a single cell can transf...versity report this week in the Proceedings of the...tified genes unique to the human egg. The identifi... functions, which may lead to solving problems fro...
Other Biology News:Fishy cooperation 2Natural anti-viral enzyme helps keep cancer cells alive, researchers find 2Scientists crack genetic secrets of human egg 2