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Luciferase assay cloning


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2 replies to this topic

#1 Azk Blub Clima

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Posted 27 December 2015 - 11:58 PM

Hello Every body,

 

I need your precious help on cloning of specific gene (in my case Enolase) into pSF-CMV-FLuc vector.

 

What kind of restriction enzyme I should use?

 

I have designed primer manually, but there specificity is in doubt. I have re-designed by Primer Blast software and i plan to use this one. Any one with better suggestion?

 

John



#2 pito

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Posted 28 December 2015 - 06:51 AM

Hello Every body,

 

I need your precious help on cloning of specific gene (in my case Enolase) into pSF-CMV-FLuc vector.

 

What kind of restriction enzyme I should use?

 

I have designed primer manually, but there specificity is in doubt. I have re-designed by Primer Blast software and i plan to use this one. Any one with better suggestion?

 

John

 

No one can answer this unless you tell them what the sequence is or more important: what restriction sites you have!

What is the MCS of that vector?

 

And you designed primers? How can you design them already if you do not know the restriction sites?


If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#3 Andrea Fortina

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Posted 20 June 2016 - 07:00 PM

Could you provide your gene sequence? It will be helpful for us to analysis.


Andrea Fortina

Genius only means hard-working for all one's life.

CreativeBiomart, Recombinant protein expert

 

 





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