HomeFAQsWhy are there high molecular weight smears or DNA in the wells of an agarose gel after a PCR using Phusion Hot Start Flex DNA Polymerase?
FAQ: Why are there high molecular weight smears or DNA in the wells of an agarose gel after a PCR using Phusion Hot Start Flex DNA Polymerase?
Strong protein binding to the DNA may be preventing proper gel separation. Try adding 1% SDS to the loading dye just before running the gel. Or non-specific products are being amplified. To avoid this we suggest the following:
Reduce polymerase concentration.
Shorten extension time.
Reduce total number of cycles.
Increase annealing temperature or try 2-step protocol.
Optimize Mg2+ concentration.
Lower primer concentration.
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.
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.