pCMV-SPORT6 empty vector

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roxanalibe
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pCMV-SPORT6 empty vector

Post by roxanalibe » Mon Jul 26, 2010 9:17 am

Hi,

I need to obtain the empty vector pCMV-SPORT6, for control, and I never obtained an empty vector out of an one with an insert. Any idea how I could do that (tips for a better ligation of the empty vector)? the mcs has: ecorI, xmaI, smaI, ecorV, salI, notI,xbaI, xhoI...the insert is cloned with salI and NotI.

Thank you for your input

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JackBean
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Post by JackBean » Mon Jul 26, 2010 12:56 pm

buy it from the provider?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

roxanalibe
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Post by roxanalibe » Mon Jul 26, 2010 2:21 pm

in this way i will never learn how to do it :-)

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JackBean
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Post by JackBean » Mon Jul 26, 2010 5:40 pm

aah, sorry, I wrote it wrong, that you do NOT want to advice to get it from vector with insert :-D

Well, I would not suggest to cut it with some RE in the sides, because then you would loose the MCS and the vector would be useless ;) Anyway, you can simply cut off your your insert with your restriction enzymes, then just make your ends blunt and ligate ;)
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

Rquantin
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Post by Rquantin » Wed Aug 04, 2010 4:20 am

Hi Roxanalibe,

I successfully get my insert out of the vector but unfortunately for whatever reason the vector will not ligate onto itself. In case you manage to do it would be possible to send me some of the empty pCMV-sport6? I would appreciate a lot.

Thanks

Rquantin

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