High Molecular Weight Material Stuck in Wells (End-point PCR Primers)

Question

Accepted Answer

Ethidium bromide-stainable material remains in the gel wells and does not migrate during electrophoresis Common causes: 1) Too many PCR cycles performed 2) Excessive DNA added to reaction 3) Formation of high molecular weight aggregates or concatenated products Solutions: 1) Reduce number of PCR cycles 2) Decrease amount of template DNA added to reaction 3) Heat sample to 65°C before loading 4) Place sample on ice immediately after heating and before loading onto gel