Can anyone help me understand what do different bands mean in gel electrophoresis after performing PCR?
If the bands fall into 2 or 3 sizes then you are possibly amplifying a polymorphic repeat sequence but if they are many bands as well as your expected band then you are probably using too low an annealing temperature so the primers are annealing at many places in the genome and producing many bands. In this case raising the annealing temperature often makes the expected band stronger while the wrong sized bands vanish with increasing temperature
Paul Rutland Thank you for responding. What does it mean if there is a faint band visible near 10,000 bp thought the expected band should be near 700bp
It probably means that there is a small amount of non specific priming in your pcr. To minimise it you can use less dna, less primer and make your annealing and extension times shorter in your pcr reaction, If these do not work then adding 5% DMSO to the reaction mix often works
- Badges
- Science method