Hi ! You can try to buy GFP Plasmids from OZ Biosciences. I'm using their PVectOZ (GFP and LacZ) Plasmid for transfection control in HUVEC. it works really well !!

Hi ! You can try to buy GFP Plasmids from OZ Biosciences. I'm using their PVectOZ (GFP and LacZ) Plasmid for transfection control in HUVEC. it works really well !!

you can ind with all big companies like sigma, open biosystems etc they have a long range. one suggestion is to be sure that for what purpose and what kind of method of transfection you want to do. e.g lentiviral or lipofectamine etc.

Hope on those two site you find the best you want.

best o luck

prasoon

CLontech provides a large array of vectors, and those based on enhanced GFP work well for me. Yo ucaqn choose different ones for targetting to organelles if that seems appropriate, and different colors if you have another fluorescent detection method you are using in the same cells for another purpose. The vectors look pretty expensive these days, because they have so many feature, some of which allow for complex experiemnts. The standard enahanced GFP from years ago woreks fine, anf putting a promoter in the MCS of pEGFP-1 plasmid gives good results in mammalian cells. I have a plasmid like that I use a lot, but am not sure if there are problems in international shipping of that type of "dangerous recombinant DNA" that might make sending it to you problematic.

There is a company in Cambridge MA, called AddGene that have a great number of plasminds contact Lianna Swanston email addess is [email protected]

I agree with Michael Bradbury, you can get GFP tagged vectors from clonetech. While choosing vectors please look for N-terminal or C-terminal tags, based on your application.

If you don't want to spend money and have some time, you can ask in the lab of Roger Tsien. You just have to sign an MTA and they will send you an aliquot. Of course you would have to pay for shipping. If you find someone who has Tsien plasmids, you only need the MTA of the Tsien lab and then that third person is allowed to send you that plasmid. I would do it, but I only have plasmids under the Tsien MTA that to not express GFP unconditionally.

Nobody who has commercial plasmids is allowed to give you an aliquot, this is regulated in the purchase conditions. It is usually only allowed to exchange proprietary plasmids within the same institution. Is there nobody in your university who has a GFP plasmid?

If you want to purchase a vector for transient transfection in mammalian cells, I suggest (as others did before) a eGFP variant, preferrably with a CMV promoter. If you have the possibility to get a monomeric mutant of GFP like the Clontech AcGFP I would choose this one over the classical/enhanced GFP, since the monomeric variants could later be used as fusion tags and would provide better results in that setting.

Good luck

In my institute I found pGLO and when I searched I found that this encodes GFP for bacterial expression and basically for teaching purposes.

Abdulmonem,

you cannot use this plasmid for transfection of eukaryotic cells, the promoter is for bacteria... You could subclone the GFP insert into a mammalian expression vector, but I am not shure whether it will actually work well enough for your application since many available and good expressing GFP variants are optimized for mammalian codon usage.

Addgene can be the answer ;)

If you only want to use the GFP-plasmid as a transfection control (so you don't mind whether it's expressed in a specific organelle/localisation), I would look in Addgene.

Addgene is a plasmid repository, usually much cheaper than commercial ones. You buy it and you get it on the mail in a couple of weeks. I've never had any issues with them.

An example:

pcDNA3-EGFP

http://www.addgene.org/13031/

Addgene is a source for many plasmids including eGFP tagged vectors with resealable price about 65$

I have used GFP-tagged vectors from Origene with good success!

Addgene,clonetech,origene,sangers, promega are the good choices...

Thanks all to help me in this topic. I got the information i need from your discussion. Special thanks to Dr. Mechael for his offer of a GFP plasmid. Also special thanks to recommend Addgene, i think it is a good resource.

FYI..Addgene just added hundreds of new vectors from Fluorescent protein fusions to the collection.  See http://www.addgene.org/fluorescent-proteins/

Thanks Joanne, Last month I got a collection of 6 fluorescent plasmids from Addgene. It is really a great organization. 

Just make sure you get a vector which expresses in your cell line. Some of vectors are just cloning vectors, and will only express in something like ecoli. For mammalian expression, I typically use pMaxGFP from Lonza because it comes as a positive control in their electroporation kits. If I need more of it, I'll just clone it into ecoli, and prep however much I need in the foreseeable future.

Good points Martin,

FYI...Addgene has many excellent vectors and can also provide positive controls.  If ever in doubt, call Addgene or email us for tech help.  One of our PhD molecular biologists can help you find what you need.

Thanks for the advice, I obtained pcDNA3-EGFP, pcDNA3-EYFP, pcDNA3-ECFP and pcDNA3-mRFP. I intend to express them in mammalian cells. Currently I amplified and extracted using maxi prep kit and I successfully got the high purity plasmids (about 500 ug each). Next step I will try the transfection. I assume they can transfect mammalian cells, can't?

Hi Abd,

It will depend on your cell type. Electroporation is typically the most efficient way for a wide variety of cells. The only other issue could potentially be the size of the plasmid, but most are typically 3-5kb so that should be fine.

M

Thanks Martin