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What DCH5a cells are? / pCAMBIA1301 isolation from DHC5a cells

pCAMBIA1301 isolation purification vector DHC5a

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#1 Curvularia protuberata

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Posted 13 January 2015 - 06:02 PM

Hey guys, I'm new here, 

well, I am about to isolate pCAMBIA1301 vector from DHC5a cells, in order to carry on with the procedure that requires multiple digestion of that vector.

The problem is that I can not find in what specie DHC5a cells belong (for instance whether they are E. coli or other specie). As a result, I don' t know what protocol to follow. The kit I am going to use has a section about isolation from E.coli and a different section about gram+ cells. I googled "DHC5a cells" , but a result I found was about a sea urchin's gene. So I don't know if I can trust this result.

Does anybody work with those kind of cells and can explain to me any tips about how to isolate plasmids from them or whether they need special treatment compaired to E. coli?

Thanks in advance :) 



#2 bob1

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Posted 13 January 2015 - 11:46 PM

Are you sure you aren't using DH5alpha cells? These are a very commonly used strain of E coli for propagation of plasmids.



#3 CPRES

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Posted 14 January 2015 - 07:21 AM

You can call up the company/lab where you got that isolate from. pCAMBIA1301 vector is grown in e.coli (as bob1 said) or agrobacterium (other gram negative rods in soil), but I could not find these cells. More likely, it does not matter. Just grow some, keep a backup glycerol stock, and do your miniprep.

 

Godspeed. We will await your update on the matter.


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#4 Curvularia protuberata

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Posted 15 January 2015 - 08:53 AM

Thanks a lot guys!! :D Your answers were really helpfull. They are E. coli indeed, I asked my supervisor today, so it's dublechecked. Thanks again :) 

 







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