Navigation Links
Efficient Delivery of siRNAs to Human ,,, Primary Cells: Electroporation ,,, vs. Chemical Transfection

expression was calculated as a percentage of gene expression compared to the negative control siRNA. Duplicates were performed for each sample.


Electroporation
An alternative to chemical transfection-mediated nucleic acid delivery is electroporation. This method involves applying an electric field pulse to induce the formation of microscopic pores (electropores) in the cell membrane. This allows molecules, ions, and water to traverse the membrane. Under specific pulse conditions, the electropores reseal and the "electroporated" cells recover and continue to grow. A distinct advantage of electroporation is that it is not dependent on cell division, and reduction in gene expression can be detected a few hours after nucleic acid delivery. This is important when working with primary cells since most divide relatively slowly if at all.

We used electroporation (Nucleofector, Amaxa) to deliver Cy3-labeled siRNA targeting GAPDH into HUVEC cells. A HUVEC-specific protocol (Amaxa) was used. We were pleasantly surprised to find that almost every cell had taken up detectable amounts of Cy3-labeled siRNA (Figure 2). The Nucleofector was then used to electroporate HUVEC, NHEK, and normal human dermal fibroblasts neonatal (NHDF-Neo) with five different concentrations of GAPDH siRNA. Real time RT-PCR revealed a dose dependent reduction in GAPDH expression in all three cell types. Some toxicity was observed at the highest siRNA dose tested (2.66 g), but there was a complete absence of non-specific effects at 1.33 g siRNA (Figure 3). To ex
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Efficiently Insert Unique Restriction Sites into Plasmid Vectors
2. Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
3. Mammalian Expression Vector for Efficient Cloning of PCR Fragments
4. Efficient Transfection of Neurospora Crassa
5. Efficient Transfection of Neurospora Crassa
6. Efficient Recovery of Ultrapure Plasmid DNA
7. Efficient and Reliable PCR Setup Using Eppendorf MasterMix
8. Efficient DNA transfection of primary CNS neurons using TransMessenger Transfection Reagent
9. Efficient RNAi-mediated gene silencing in neuronal cells using QIAGEN siRNA and TransMessenger Transfection Reagent*
10. Cloning Based on Efficient Three-Fragment Assembly DNA Ligation
11. Efficient and Reliable Linear Amplification of cRNA
Post Your Comments:
(Date:9/2/2015)... ... September 02, 2015 , ... The ... the use of special anti-vascular endothelial growth factor agents given by injection into ... does not have a scientific protocol for the many millions diagnosed with it. ...
(Date:9/2/2015)... Sep. 02, 2015 ... has announced the addition of Jain PharmaBiotech,s ... and Companies" to their offering. ... technologies besides fluorescence in situ hybridization (FISH), ... Molecular cytogenetics includes application of nanobiotechnology, microarrays, ...
(Date:9/2/2015)... CUMBERLAND, R.I. , Sept. 2, 2015 /PRNewswire/ ... has been enrolled in the multicenter Phase 2 ... for the long-term treatment of recurrent subfoveal choroidal ... AMD).  NT-503 is a unique vascular endothelial growth ... versatile ECT implant. "This landmark proof-of-concept ...
(Date:9/2/2015)... Calif. , Sept. 2, 2015 Veracyte, ... Bonnie H. Anderson , president and chief executive officer, ... on Wednesday, September 16, 2015 at 10:30 a.m. ET ... The live audio webcast and subsequent replay ... The webcast will be available shortly after conclusion of ...
Breaking Biology Technology:StemCell GeneticMed Announces That It Has Started Stem Cell Treatments for Age Related Dry Macular Degeneration (AMG) 2Global Cytogenetics Technologies Study 2015-2025 - Latest Markets and Companies Analysis 2Global Cytogenetics Technologies Study 2015-2025 - Latest Markets and Companies Analysis 3Neurotech Announces First Patient Enrolled in Novel Anti-VEGF Encapsulated Cell Therapy Study 2Neurotech Announces First Patient Enrolled in Novel Anti-VEGF Encapsulated Cell Therapy Study 3Neurotech Announces First Patient Enrolled in Novel Anti-VEGF Encapsulated Cell Therapy Study 4Veracyte to Present at Morgan Stanley Global Healthcare Conference 2
... to the constant whir of a computer's fan or held a ... , ,In fact, today's ultra-powerful computers generate so much heat ... mechanical engineering professor Tim Shedd . , ,Shedd ... method that early tests show can remove heat at rates up ...
... Wis. - The typical weekend bicycle rider might ... purchase. But for the professionals of the cycling world, it's ... , , Trek Bikes of Waterloo, Wis., operates under ... available technology for Lance Armstrong as he seeks a seventh ...
... the use of e-mail on vacation or the weekend? According ... email while on vacation. Most people check their email an average ... one mail address and a quarter of the respondents, said they ... , ,Original air date: Saturday, July 9, 2005 , , ...
Cached Biology Technology:A new tool for keeping computers cool 2A new tool for keeping computers cool 3A high-tech Trek at the Tour de France 2A high-tech Trek at the Tour de France 3
(Date:8/6/2015)... JOSE, Calif. , Aug. 6, 2015 /PRNewswire/ ... leading developer of human interface solutions, today announced ... TouchPad technologies on Windows 10. Microsoft leveraged Synaptics, ... to the operating system. Through ... drivers are fully certified with Microsoft,s Precision TouchPad ...
(Date:8/4/2015)... , Aug. 04, 2015 ... http://www.researchandmarkets.com/research/nvh7k2/digital_forensics ) has announced the addition of ... (Hardware, Software, and Service), Sub-Segment (Computer Forensics, ... Forensics), Tool Types, Service, Vertical and Region ... to their offering. By Component ...
(Date:7/31/2015)... de 2015 BGI llevará a cabo la 10ª ... al 25 de octubre de 2015, en ... su décimo aniversario este año. Desde su inauguración en ... reuniones anuales más influyentes del mundo en los campos ... científicas más dinámicas, entusiastas y amenas. ICG-10 ...
Breaking Biology News(10 mins):Synaptics Chosen for Human Interface Co-Development on Windows 10 2Digital Forensics Market - The Market Represents a Compound Annual Growth Rate (CAGR) of 11.3% From 2015 to 2020 2La 10ª Conferencia Internacional sobre Genómica (ICG-10) comienza en octubre 2La 10ª Conferencia Internacional sobre Genómica (ICG-10) comienza en octubre 3
... and computer programming equally. So when Rios who ... of Waterloo went looking for a graduate program, ... at Florida State University, which has about 35 graduate ... sciences in fall 2010. Now, along with his ...
... has elected ten new Fellows of the Society for ... in one or more of the following: research, teaching, ... during Entomology 2011 -- ESA,s 59th Annual Meeting -- ... Nevada (see: http://www.entsoc.org/entomology2011 ). Dr. Susan ...
... Neurotechnology , a provider of high-precision biometric identification ... new voice identification technology. VeriSpeak is now available ... as a new voice identification component in the ... VeriSpeak combines voiceprint identification and phrase recognition technologies, ...
Cached Biology News:New software aids fight against nitrates in Florida's groundwater 2New software aids fight against nitrates in Florida's groundwater 3Entomological Society of America names 2011 fellows 2Entomological Society of America names 2011 fellows 3Entomological Society of America names 2011 fellows 4Entomological Society of America names 2011 fellows 5Entomological Society of America names 2011 fellows 6Entomological Society of America names 2011 fellows 7Entomological Society of America names 2011 fellows 8Entomological Society of America names 2011 fellows 9Entomological Society of America names 2011 fellows 10Entomological Society of America names 2011 fellows 11Entomological Society of America names 2011 fellows 12Neurotechnology Introduces VeriSpeak Voice Identification Technology 2Neurotechnology Introduces VeriSpeak Voice Identification Technology 3Neurotechnology Introduces VeriSpeak Voice Identification Technology 4
The Programmable MP-5000P-F Power Supply can be applied for any Microfluidic application....
Rab GDP dissociation inhibitor beta...
... POWER 500V Programmable Power Supply are ... can handle virtually any application from ... blotting. It is unique in offering ... MAX POWER Programmable Power Supply is ...
Handy 100V Power Supply is Low-cost, compact and lightweight. Easy to operate - either 50 or 100 VDC is selectable as the output voltage. These voltages are sufficient for ordinary electrophoresis....
Biology Products: