Home Resources Video Library Quick Tips - Why do I have so many wildtype colonies?

Quick Tips - Why do I have so many wildtype colonies?

This video explains why you might have a large number of wildtype colonies following site-directed mutagenesis and transformation.

Script

Customers often ask, “Why is the background so high? Why do I have so many wildtype colonies?”

Wildtype colonies result from template DNA that is carried over from the PCR into the final transformation step. To eliminate template DNA, first, be sure that the PCR reaction has worked efficiently. Then, use only 1ul of the PCR reaction in the KLD reaction. This will eliminate plasmid DNA and reduce the number of background colonies.

If you have other questions you can always contact us at info@neb.com.

Related Videos

  • TroubleshootingTipsQ5SiteDirectedMutagenisis_720
    Troubleshooting tips for Q5® Site-Directed Mutagenesis Kit
  • NEBTV_DNA-Assembly-Cloning-and-Mutagenesis-Kits_1920
    Quick Tips - Can I make multiple mutations with the Q5® site-directed mutagenesis kit?
  • OverviewOfQ5SDMKit_720
    Overview of the Q5® Site-Directed Mutagenesis Kit
Visit NEB’s Video Library

Choose your country

North America

Europe

Asia-Pacific

If you don't see your country above, please visit our international site

Session Expired

You have been idle for more than 20 minutes, for your security you have been logged out. Please sign back in to continue your session.

Institution Changed

Your profile has been mapped to an Institution, please sign back for your profile updates to be completed.

Sign in to your NEB account

To save your cart and view previous orders, sign in to your NEB account. Adding products to your cart without being signed in will result in a loss of your cart when you do sign in or leave the site.