Hi All,
It will be great if somebody can tell me the source for mammalian expression plasmids with tags.
Looking forward to hear from you.
Regards,
Niraj
0
Mammalian expression vectors with HA, c-myc, FLAG or anyother tag
Started by nirajm, Jun 12 2009 04:10 PM
3 replies to this topic
#2
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Posted 15 June 2009 - 07:35 AM
nirajm, on Jun 12 2009, 08:10 PM, said:
Hi All,
It will be great if somebody can tell me the source for mammalian expression plasmids with tags.
Looking forward to hear from you.
Regards,
Niraj
It will be great if somebody can tell me the source for mammalian expression plasmids with tags.
Looking forward to hear from you.
Regards,
Niraj
Try Addgene (http://www.addgene.org/pgvec1). It is a share site that provides plasmids submitted from academic groups in the same way that you request vectors from other labs. You can obtain numerous vectors for about $65 ea. if you sign an MTA.
Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley
#3
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Hmmm, I think it's working
Posted 22 July 2009 - 04:33 PM
There is also http://plasmid.med.harvard.edu/PLASMID/Ord...erview.jsp#cost for about $45/clone.
Also if you have an empty vector and you want to add a tag, just add the tag to a PCR primer for a target gene and amplify from there. If you add a restriction site on either side of the tag, you should be able to chop it out and re-liagte it into other plasmids too.
Also if you have an empty vector and you want to add a tag, just add the tag to a PCR primer for a target gene and amplify from there. If you add a restriction site on either side of the tag, you should be able to chop it out and re-liagte it into other plasmids too.
#4
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Posted 22 July 2009 - 04:38 PM
bob1, on Jul 22 2009, 07:33 PM, said:
There is also http://plasmid.med.harvard.edu/PLASMID/Ord...erview.jsp#cost for about $45/clone.
Also if you have an empty vector and you want to add a tag, just add the tag to a PCR primer for a target gene and amplify from there. If you add a restriction site on either side of the tag, you should be able to chop it out and re-liagte it into other plasmids too.
Also if you have an empty vector and you want to add a tag, just add the tag to a PCR primer for a target gene and amplify from there. If you add a restriction site on either side of the tag, you should be able to chop it out and re-liagte it into other plasmids too.
I just did this myself using FLAG. Added it to pBluescript, and then added a restriction site in front of the tag, so I can make C-terminal FLAG constructs. Can also insert flanking sequence downstream of the tag if I ever want to express the tagged protein as a single copy in the genome as opposed to plasmid-borne. Went together pretty easily. It required some polymerase, restriction enzymes, and some sequencing. All in all, likely cheaper than what I would've paid (was looking at Sigma for FLAG vectors), and probably more versatile with being able to add flanking sequence behind the tag.
As for places that sell vectors, I remember seeing many at Sigma for FLAG. Almost any supplier of molecular biology supplies probably has a good number of tag vectors to choose from. But as stated above, you'll probably pay at least 5 or 6 times more than if you'd get one from the links above.
Edited by fishdoc, 22 July 2009 - 04:43 PM.
