Protocol Online logo
Top : Forum Archives: : Molecular Cloning

RE of PCR Fragment - (Mar/17/2008 )

Hi there, I am trying to digest a PCR frament with two different RE NdeI and XhoI in sites I have added to the end. Does anyone know how many extra nucleoties are requirewd for these two enzymes to cu properly?. Where can I find the same information for all the enzymes? I have beedn trying to do a ligation for a long long times without sucess and I be,ive the reason is becouse I only added 6 nucletides and I need more, at least for XhoI. Thank you

-maira-

QUOTE (maira @ Mar 17 2008, 05:27 PM)
Hi there, I am trying to digest a PCR frament with two different RE NdeI and XhoI in sites I have added to the end. Does anyone know how many extra nucleoties are requirewd for these two enzymes to cu properly?. Where can I find the same information for all the enzymes? I have beedn trying to do a ligation for a long long times without sucess and I be,ive the reason is becouse I only added 6 nucletides and I need more, at least for XhoI. Thank you


It depends on your enzyme, have a look here: http://www.neb.com/nebecomm/tech_reference...nucleotides.asp

-hobglobin-

thanks. Tom

-maira-