We decided to revisit a popular blog from our Promega Connections past for those of you in the amplification world. Enjoy:

• • Modify reaction buffer composition to adjust pH and salt concentration.
  • Titrate the amount of DNA polymerase.
  • Add PCR enhancers such as BSA, betaine, DMSO, nonionic detergents, formamide or (NH₄)₂SO₄.
  • Switch to hot-start PCR.
  • Optimize cycle number and cycling parameters, including denaturation and extension times.
  • Choose PCR primer sequences wisely.
  • Determine optimal DNA template quantity.
  • Clean up your DNA template to remove PCR inhibitors.
  • Determine the optimal annealing temperature of your PCR primer pair.

[Drum roll please]…and the  most important thing you can do to improve your PCR results is:

• Titrate the magnesium concentration.