This is a detailed troubleshooting tip for PCR.
Why do I have non-specific bands when I run my gel?
Causes：Template concentration is inappropriate
Measures：Use appropriate template concentrations. For a 50 μl
PCR reaction recommended concentrations are:
human genomic DNA= 0.1-1 μg; E.coli genomic
DNA = 10-100ng; lphage DNA= 0.5-2.5 ng;
plasmid DNA 10-100 ng
Causes：Damaged template DNA
Measures：Minimize damage to template DNA by avoiding vortexing,
heat treatment, strong UV, shearing or ultra sonication
Causes：Denaturation time is too short
Measures：Optimize the denaturation time in increments of 5 seconds
Causes：Denaturation temperature is too low
Measures：Optimize the temperature in increments of 0.5°C
Causes：Annealing temperature is too low
Measures：Raise the temperature in increments of 2°C
Causes：Extension time is too short
Measures：Lengthen the extension time in increments of 1 minute
Causes：Cycle number is too high
Measures：Reduce the number of cycles in decrements of 2 cycles