Where to buy Vectors

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mehdi71000
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Where to buy Vectors

Post by mehdi71000 » Thu May 24, 2007 2:25 pm

Hi

I’ve been looking for a wile on the net for some GFP /BFP /YFP / RFP and plant growth hormone Vectors. To use on plants and small animals but no luck. Can any one direct me to a suitable distributor please much appreciated :D

david23
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what stage are you at now?

Post by david23 » Mon May 28, 2007 8:12 am

The websites you showed me did have the plasmids, but they seemed a little expensive didnt they?

mehdi71000
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Post by mehdi71000 » Mon May 28, 2007 11:03 am

yes i found on it Gwiz GFP. its around 180£ for 1 mg.

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Post by david23 » Tue May 29, 2007 1:09 am

thats more than enough for many many experiments. But to prevent wasting all, u should really clone it in bacteria first. if u need help with it, then buy a really really cheap DNA sequence. which are sold a couple cent/base pair, buy some random sequence or one with repeating AAAA to test first.

mehdi71000
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Post by mehdi71000 » Tue May 29, 2007 10:31 am

So for the experiment itself do I need to attach it to another vector before injection or its fine as it is? gWiz™ High-Expression Luciferase vector (gWiz™ Luciferase). Could for cloning i could just inject it into a cells nucleus and after it has multiplied. I could crush the cells and by electropolis separate the GFP?
or for the expression of gfp I need to attach the vector to another vector? :?

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Post by david23 » Wed May 30, 2007 6:10 am

the GFP should be sold as part of a vector/plasmid, since thats where GFP is used the most, for genetic experiments. the luciferase vector does the same job as a GFP vector. Clone it in bacteria, because bacteria colonies are easier to culture and identify on a petri plate. With fish embryos, or any these complex organisms, the culturing process is difficult. Also bacteria genomes are simpler, and contain fewer genes, so identifying the GFP plasmid on a gel electrophoresis is easier. When you clone it in bacteria, you are eventually trying to extract the GFP plasmid from it. The GFP and its plasmid should always be together.

The luciferase vectors are more expensive right?

mehdi71000
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Post by mehdi71000 » Wed May 30, 2007 12:32 pm

That’s all they had? luciferase vectors. But this GWIZ is very bright in the pictures I searched. Is this vector just the gfp but it expresses the gene more than usual?or they designed a whole new protein?

Thanks for your help. You’re a very smart man?

mehdi71000
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Post by mehdi71000 » Fri Jun 01, 2007 12:07 am

is gfp and luciferase the same thing or one is brighter than other?

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canalon
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Post by canalon » Fri Jun 01, 2007 2:25 am

Gfp is fluorescent, you need to excite it with another light with a shorter wavelength to see it glow. On the other hand luciferase is chemiluminescence, it needs a substrate and create light. Usually plasmids includes a whole lux operon which will provide the pathway for the substrate synthesis and the luciferase to make it glow.
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

mehdi71000
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Post by mehdi71000 » Sat Jun 02, 2007 12:10 am

thanks guys.
thanks canalon for the heads up on gfp i totally had the wrong idea about it.your a life saver

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