Troubleshooting/Ligation

Troubleshooting

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• General Tips and Tricks
1. Transformation
2. Ligation and Restriction Digest
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Page is under construction.
iGEM HQ is currently working on updating this information for the iGEM 2015 competition.

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 and restriction digests.

No colonies

If you know your cells are working well, there are a few common ligation problems that might be happening with your reaction.

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

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