How do I increase the intensity of PCR bands?

If you are experiencing faint or no bands, try the following tips to increase band intensity:

• Use the appropriate number of PCR cycles, usually the number of cycles is carried out 25-35 times, but in cases where the template concentration is low use more cycles
• Optimize the extension time for successful replication of the target, for most PCR applications an extension time of 1 min/kb is sufficient
• Make sure there is sufficient annealing time for primers to bind to the template
• Optimize annealing temperature, primers cannot bind to the template if the temperature is too high
• Use a denaturation temperature of 95 °C
• Optimize the denaturation time, if the denaturation time is too long or too short, then the DNA might degrade or not completely denature, respectively
• Each dNTP should have a concentration of 200 µM
• Optimize your primer concentration, the concentration of each primer should range between 0.1 to 0.5 µM, but for most PCR applications 0.2 µM is sufficient
• Avoid cross-contamination, contaminated dNTPS and impure water, always use fresh nuclease-free water
• Make sure to use sufficient amounts of magnesium, for PCR a typical final concentration of magnesium ranges from 1 to 4 mM