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miRNA 3´UTR target expression vectors


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6 replies to this topic

#1 Nádia.c

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Posted 19 July 2010 - 10:18 AM

Hello everyone

I want to clone the UTR of my gene of interest in one expression vector, downstream of luciferase or GFP gene to access if it's controlled by miRs. Since after a lot of attempts I couldn’t clone my UTR my boss opened the wallet and said that I could by an already made vector with my UTR!!!

So my question is, does anyone know companies that sell 3´UTR target expression vectors? I only know switchGear and Genecopoeia, but my UTR is not in their list.

Do you know any other company?

tks

Best regards

#2 ljchen

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Posted 20 July 2010 - 01:13 AM

ABI: pMIR

Promega:pmirGLO Dual-Luciferase miRNA Target Expression Vector ...............In fact, I prefer this one.




Hello everyone

I want to clone the UTR of my gene of interest in one expression vector, downstream of luciferase or GFP gene to access if it's controlled by miRs. Since after a lot of attempts I couldn’t clone my UTR my boss opened the wallet and said that I could by an already made vector with my UTR!!!

So my question is, does anyone know companies that sell 3´UTR target expression vectors? I only know switchGear and Genecopoeia, but my UTR is not in their list.

Do you know any other company?

tks

Best regards



#3 Nádia.c

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Posted 26 July 2010 - 02:45 AM

Hi

thanks for your answer, but I know those two vestors, but I want companies that sells the UTR already cloned.... do you know any?

#4 KevinK

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Posted 12 August 2010 - 05:52 AM

Nadia,
I would contact SwitchGear as they should have all of the 3'-UTRs available. If they have not made it already my understanding is that it will be made for the order.

As was mentioned, Promega does make the pmirGLO vector, but requires cloning. The advantages of this vector are that it has a Renilla luciferase gene for normalizaiotn of transfection efficiency, and once you make it you have an unlimited supply of the plasmid. Many studies eventually involve making mutations of miRNA binding sites or other elements, so you might be better off making the construct in the first place.

Please don't hesitate to contact Promega Technical Services if you have questions or for troubleshooting assistance with the cloning. I know how frustrating it can be when the cloning gods are not on your side, but they evenually come around.

Regards,
Kevin
Promega Corporation
Madison, WI

#5 Nádia.c

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Posted 31 August 2010 - 06:40 AM

Hi Kevin

Tks for your advice.I contacted SwitchGear and they are not planning to clone my UTR of interest :(
Promega has a partnership with Switchgear to make a UTR library in pmirGLO vector, am I right? I suppose this includes only the most popular genes...

I really want to avoid the clonning since I tried for long to clone the UTR in the pGL3 and has been a nightmare. I don't want to wast more time with this. My UTR is 3 kb long so It's hard...


Nadia,
I would contact SwitchGear as they should have all of the 3'-UTRs available. If they have not made it already my understanding is that it will be made for the order.

As was mentioned, Promega does make the pmirGLO vector, but requires cloning. The advantages of this vector are that it has a Renilla luciferase gene for normalizaiotn of transfection efficiency, and once you make it you have an unlimited supply of the plasmid. Many studies eventually involve making mutations of miRNA binding sites or other elements, so you might be better off making the construct in the first place.

Please don't hesitate to contact Promega Technical Services if you have questions or for troubleshooting assistance with the cloning. I know how frustrating it can be when the cloning gods are not on your side, but they evenually come around.

Regards,
Kevin



#6 Earnest

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Posted 28 March 2011 - 08:18 AM

Hi Kevin

Tks for your advice.I contacted SwitchGear and they are not planning to clone my UTR of interest :(
Promega has a partnership with Switchgear to make a UTR library in pmirGLO vector, am I right? I suppose this includes only the most popular genes...

I really want to avoid the clonning since I tried for long to clone the UTR in the pGL3 and has been a nightmare. I don't want to wast more time with this. My UTR is 3 kb long so It's hard...



Nadia,
I would contact SwitchGear as they should have all of the 3'-UTRs available. If they have not made it already my understanding is that it will be made for the order.

As was mentioned, Promega does make the pmirGLO vector, but requires cloning. The advantages of this vector are that it has a Renilla luciferase gene for normalizaiotn of transfection efficiency, and once you make it you have an unlimited supply of the plasmid. Many studies eventually involve making mutations of miRNA binding sites or other elements, so you might be better off making the construct in the first place.

Please don't hesitate to contact Promega Technical Services if you have questions or for troubleshooting assistance with the cloning. I know how frustrating it can be when the cloning gods are not on your side, but they evenually come around.

Regards,
Kevin


I know this seems late, but I only saw this now. These are in stock and popular. They have a massive sale in miRNA stuff now. Have a look!: http://www.abmgood.c...11838&dsn=12288

Edited by Earnest, 28 March 2011 - 08:55 AM.


#7 Nádia.c

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Posted 04 April 2011 - 06:32 AM

Thank you! I have my plasmid already, by genecopoeia. But is good to know this! thank you a lot




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