Transfection Efficiency

Transfection Efficiency

Postby postdoc on Mon Oct 26, 2009 9:15 am

Generally, transfection can be considered as a major laboratory method in integrating protein, RNA, and DNA molecules into tissues and cells. Delivery of molecules encoding genes like plasmid DNA, messenger RNA and small RNA molecules regulating gene expression (microRNA and siRNA)into the primary cells and cancer cell lines have been applied by the scientists.

Transfection efficiencies greatly depends on specific on the type of the cell, reagent used and the protocol applied. But, there is no single transfection reagent that fits different types of cells - scientists have to test multiple reagents and optimize transfection conditions in order to achieve acceptable transfection efficiency withouit inducing cellular cytotoxicity. There are several pre-optimized transfection reagents commercially available from Altogen Biosystems (http://www.altogen.com) for such cell lines as: MEF, A549, HepG2, Fibroblast, VERO, and many others. Also, in vivo transfection kits allow delivery of protein, DNA, and RNA molecules into animals (http://www.altogen.com/mirna.php).
postdoc
 
Posts: 44
Joined: Mon Oct 26, 2009 8:45 am

Re: Transfection Efficiency

Postby biologist on Mon Oct 26, 2009 12:37 pm

Chemical transfection, reverse transfection, stable transfection (and efficiecy of such transfection) is very important in the biological research and study of genomics, stem cell research, as well as development of new medicines and treatment protocols. Also, successful gene silencing experiments rely on high transfection efficiency. Choosing the right method of transfection as well as the correct transfection agent is at the basis for the success of the efficiency of various transfection experiments.

Transfections may be performed utilizing lipids, polymers, liposomes, nanoparticles, and other types of reagents. Scientists and researchers have determined that a combination of appropriate transfection reagents as well as quality of plasmid DNA (or RNA) strongly influences the results of such experiments. Using pre-optimized transfection kits and utilizing ultrapure plasmid DNA allow for optimal high transfection efficiency results.

Transfection efficiency is influenced by not only the transfection method utilized, but also the quality of DNA (or RNA) used in the process. Various parameters will affect the efficiency of any transfection attempt, including the type of transfection method, cell cultures, media/serum, etc. However, the two most important parameters for successful transfection efficiency are the quality/purity of DNA (or RNA) and the type of transfection reagent used.
biologist
 
Posts: 44
Joined: Sat Aug 01, 2009 9:48 pm

Re: Transfection Efficiency

Postby siRNA on Sun Jan 17, 2010 8:43 pm

Altogen Labs http://www.altogenlabs.com now offers a transfection service that includes the generation of characterized and stable transfectants, the optimization of transfection protocols for particular cell line or primary culture, as well as special services and customized projects that are requested by researcher. Company experience is a big part of their transfection service, they have many years of experience and have been working with scientists in this field for a long time.
Last edited by siRNA on Tue Oct 30, 2012 10:52 am, edited 1 time in total.
siRNA
 
Posts: 56
Joined: Mon Oct 12, 2009 10:00 am

K562 (human leukemia cell line)

Postby ATGC2001 on Fri Nov 05, 2010 11:33 am

K562 are of the erythroleukemia (http://en.wikipedia.org/wiki/Erythroleukemia) type and were derived in 1970 from the pleural effusion of a 53-year-old woman with chronic myeloid leukemia (CML). It was the first human immortalized myelogenous leukemia line to be established. The cells, which produce hemoglobin, can be used as highly sensitive targets in in-vitro natural killer assays as they lack the MHC complex required to inhibit NK activity.

K562 cells have been shown to spontaneously develop characteristics similar to early-stage erythrocytes, granulocytes, and monocytes. They lack any trace of Epstein-Barr virus and other herpes viruses.

K562 carry the Philadelphia chromosome with a BCR-ABL b3-a2 fusion gene and also exhibit a second reciprocal translocation between the long arm of chromosome 15 with chromosome 17. K562 is strongly positive for immunoglobuln Fc receptors and pinocytosis. It is suggested K562 is not a B cell line and while it has some T cell properties, these are not exclusive.
ATGC2001
 
Posts: 26
Joined: Thu Oct 01, 2009 7:39 am

Re: Transfection Efficiency

Postby postdoc on Sat Nov 26, 2011 11:14 pm

Transfection efficiency can be measured by Quantitation of shRNA-, or siRNA-induced mRNA knockdown by realtime qRT-PCR, or protein expression reduction by Western Blot - commercial service avaible from Altogen Labs http://altogenlabs.com/services/rnai_services/quantitation_of_siRNA_shRNA_functional_knockdown_effect_by_qRT_PCR_and_Western_Blot
postdoc
 
Posts: 44
Joined: Mon Oct 26, 2009 8:45 am

Re: Transfection Efficiency

Postby siRNA on Tue Oct 30, 2012 10:47 am

postdoc wrote:Generally, transfection can be considered as a major laboratory method in integrating protein, RNA, and DNA molecules into tissues and cells. Delivery of molecules encoding genes like plasmid DNA, messenger RNA and small RNA molecules regulating gene expression (microRNA and siRNA)into the primary cells and cancer cell lines have been applied by the scientists.

Transfection efficiencies greatly depends on specific on the type of the cell, reagent used and the protocol applied. But, there is no single transfection reagent that fits different types of cells - scientists have to test multiple reagents and optimize transfection conditions in order to achieve acceptable transfection efficiency withouit inducing cellular cytotoxicity. There are several pre-optimized transfection reagents commercially available from Altogen Biosystems (http://www.altogen.com) for such cell lines as: MEF, A549, HepG2, Fibroblast, VERO, and many others. Also, in vivo transfection kits allow delivery of protein, DNA, and RNA molecules into animals (http://www.altogen.com/mirna.php).


CRO Contract Research Transfection Services available from Altogen Labs - www.AltogenLabs.com
siRNA
 
Posts: 56
Joined: Mon Oct 12, 2009 10:00 am

Re: Transfection Efficiency

Postby medicbio on Wed Jul 02, 2014 4:49 pm

Western Blot Services also offers qRT-PCR, the gold standard for measuring your gene knockdown: https://www.westernblotservice.com/services/#assay Also listed on their website is their expedited Wes service: https://www.westernblotservice.com/western-blot-test/
medicbio
 
Posts: 7
Joined: Tue Jun 24, 2014 2:23 pm


Return to General Topics

Who is online

Users browsing this forum: No registered users and 1 guest

cron