subcloning problems and need help..please!!

Debashish Das debashish.das at ki.se
Wed May 3 03:04:57 EST 2006


hi

i am trying to subclone a target dna from pskt to plox. the target dna has a gene of interest-
linker-gfp. the 
restriction digestion of the pskt vector releases a correct size of 2.7kb. but after inserting the 
fragment in plox, we 
see the linker missing?? on sequencing we see the sequence of gfp and the gene of interest 
but no linker. the 
linker is a small fragment of 20-30 kb. the subcloning done in XL10 and XL 1 bacteria.

please let me know as to how to over come the problem...

thanx a lot..

Deb



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