do you know a vector plasmid for cloning big pcr fragments? 15-20kb!please!!!
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#1
danielaceccarelli
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Posted 28 January 2005 - 06:45 AM
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.
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!
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
