One of the most common tech calls we get at NEB is “My ligation didn’t work” or “I didn’t get any transformants. What went wrong?”
So, actually in our experience, the issue usually doesn’t arise in the ligation step. Often there are problems upstream of ligation, such as in DNA purification or in inefficient cutting during restriction.
Cloning protocols are complicated and issues can arise at many different places so we recommend starting by using the troubleshooting guide at NEB.com to help you figure out where things might have gone wrong in your particular case.
And remember, if you have any additional questions you can always contact us via email at firstname.lastname@example.org.
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