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Which vector for cloning big PCR fragments?


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#1 danielaceccarelli

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Posted 28 January 2005 - 06:45 AM

:lol: do you know a vector plasmid for cloning big pcr fragments? 15-20kb!
please!!!

#2 george@CASE

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Posted 28 January 2005 - 12:13 PM

how did you get a PCR pdt that huge?? Is this a fusion of some sort?

You may have to use BACs for that size insert, and you have to cut it with a restriction enzyme and ligate it into the MCS of the BAC.

#3 peixumol

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Posted 30 January 2005 - 03:47 AM

ˇˇyes,BAC or cosmid can be considered.the same curiosity:how can you amplify such big DNA fragment by PCR? the tip will be quite helpful for us!
paul in NanJing China
God helps them who help themselves!

#4 danielaceccarelli

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Posted 31 January 2005 - 04:06 AM

:(
i used LATaq polymerase TAKARA inc.
they say you can amplify 35 kb. i arrived to 19kb. part of a natural plasmid in V. cholerae!thanks a lot
d




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