Resources/Troubleshooting

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Cloning Troubleshooting

Welcome to the troubleshooting hub! Below are some helpful tips and troubleshooting strategies for common cloning problems in E. coli.

Cloning is difficult. No one who has ever picked up a pipette in the lab would debate this fact. However, cloning shouldn't be impossible! This page exists to help students and teams who are struggling with cloning and other molecular techniques. Treat this page as another resource in your troubleshooting endeavors rather than the be-all, end-all resource for iGEM cloning problems. I hope this will be a helpful resource for every iGEM team!

Learn about cloning troubleshooting here!

  • Read about General Tips and Tricks with a focus on:
    • Bacterial Transformations
    • Restriction Digests and Ligations

  • Still have a cloning problem? Contact Traci and ask a question (see right box for contact info)!
  • Make sure that you also ask your instructors and more experienced students around you!

Welcome!

My name is Traci, and I'm here to help you with your cloning problems.

You can contact me:

  1. traci AT igem DOT org
  2. @Traci_H_Angelli

General Tips and Tricks

Transformation Problems

For many teams, cloning problems center around transformation problems. Here I describe some general tips that can be applied to any type of assembly method when using E. coli cells as the transformation host.

Restriction Digest and Ligation Problems

Since we provide teams with nearly 2,000 BioBricks parts, I will discuss common problems students can experience when using restriction enzymes and DNA ligation assemblies, which includes BioBricks Assembly.

iGEM Protocols

Below are the links to the various protocols that iGEM Headquarters has provided over the years. These protocols can also be found through the Parts Registry page under the "Help" and "Protocols" links in the black toolbar at the top of the page. These are another useful resource that teams should read through when they're experiencing cloning problems.

BioBrick Cloning Protocols
Other Protocols for Molecular Techniques