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pUC and pSC101 vectors in the same cell


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#1 Raygoza

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Posted 25 June 2015 - 12:29 PM

Hi, i will need to transform cells containing a pUC vector with a psc101 vector. I know that they are not incompatible. However, I'm wondering if anyone has had experience doing this and what sort of levels should I expect for the psc101 plasmid. is it what i would usually get if there was the psc101 vector only, slightly less or significantly less than usual. Thanks



#2 labtastic

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Posted 25 June 2015 - 01:21 PM

Levels of what? Miniprepped plasmid? Transformed colonies? Expressed protein?


Edited by labtastic, 25 June 2015 - 01:22 PM.


#3 Raygoza

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Posted 25 June 2015 - 03:56 PM

Levels of what? Miniprepped plasmid? Transformed colonies? Expressed protein?

Levels of psc101 miniprepped plasmid.


Edited by Raygoza, 25 June 2015 - 06:17 PM.


#4 labtastic

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Posted 26 June 2015 - 08:36 AM

Are you trying to miniprep from cells grown with both pSC101 and pUC? If so, I'm unsure what the purpose of that would be rather than miniprepping each individually.

 

But if that's what you're doing, then you should get comparable quantities of each individual vector from cells harboring both plasmids as compared to cells in which the plasmids were grown separately.

 

Generally speaking inserts don't significantly impact the level of purified plasmid compared to empty vector. The origin of replication, which controls copy number in the cell, is the biggest factor. pSC101 is a very low copy plasmid, so your yields will be low compared to other typical expression vectors (especially the pUC vector).


Edited by labtastic, 26 June 2015 - 08:38 AM.





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