Any experience in improving blunt end liga

Nick Theodorakis nicholas_theodorakis at urmc.rochester.edu
Sun Jul 1 15:25:43 EST 2001


In <20010701143012.20030.qmail at ww02.jatek.com>, fly wrote:
>
>Many thanks for your help.
>Would you please let me know:
>what does 5x buffer mean, does it contain half of what the 10x buffer has? I
>have only 10x buffer, how could i make 10x into 5x buffer?
>

If you were talking to me (it's hard to tell), I was referring to the buffer in
this documemt:

http://www.invitrogen.com/Content/Tech-Online/molecular_biology/manuals_pps/tbt4ligase.pdf

The 5X Buffer is:
250 mM Tris-Cl pH7.6
50 mM MgCl2
5 mM ATP
5 mM dithithreitol
25% (wt./vol) PEG8000

That bulletin gives some tips for blunt-end ligations.

Nick


-- 
Nick Theodorakis
nicholas_theodorakis at urmc.rochester.edu

 




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