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Cloning of sequence with PCR added sites - BstBI and BclI (Jun/27/2005 )

hi everyone.
i want to amplify a gene in order to transfert it in an other vector. I want to add Bcl I site at 3' end and BstBI site at 5' end.
I've searched through NEB and didn't find any information on capability of these enzymes to cleave close to the end.
Does anyone have made such experiment with one or both of these enzymes ? Can you give me tips about bp to add after these restriction sites?

Many thanks.


Hi Fred,

Have you read this note on NEB site:

Note: As a general rule, enzymes not listed below require 6 bases pairs on either side of their recognition site to cleave efficiently.


blink.gif oups i must admit i didn't read that...
thank you very much. I'm gonna try it and tell you the success of it.