Troubleshooting/Ligation

Troubleshooting

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• General Tips and Tricks
1. Transformation
2. Ligation
3. Restriction Digest
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Ligation Troubleshooting

Now that you know you transformation efficiency and it's above 1 x 10⁸ CFU/µg DNA, we can work on other possible problems if you're still not getting great results from your cloning. This page is focused on common problems students have with ligations.

No colonies

1. Uncut plasmid: It's possible that your destination backbone was not cut well during your digest.
2. Ligase didn't work:

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