Protocol Online logo
Top : Forum Archives: : Molecular Biology

factors contribute to the ligation failure.. - (Jun/06/2005 )

hye all..

i wanna know what are the common factors that usually made the ligation (cohesive ends)process unsuccessful?and do we need to put ATP in our ligation mixture?do we really need to put PEG too? curiosity could kill me know tongue.gif


No, you don't need to add ATP because ligation buffer contains ATP and that's why repeated freeze and thawing of the buffer should be avoided. Sometimes the problem is not with the ligation itself but lies at vector and insert preparation.