Luciferase assay cloning - (Dec/28/2015 )

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

Azk Blub Clima on Mon Dec 28 07:58:44 2015 said:

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?

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