Troubleshooting/Ligation

Troubleshooting

• Introduction
• General Tips and Tricks
1. Transformation
2. Ligation
3. Restriction Digest
• iGEM Protocols
• Answers
• Ask Your Questions

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

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:

• Press Kit
• Newsletter

• Upload Files
• Recent Changes
• Special Pages

• Requirements
• Calendar
• Meetups
• Giant Jamboree

• About
• Contact
• Sponsor
• Registry
• igem.org