- Be sure to use an annealing temperature determined by the nearest neighbor method (or Tm calculator).
- Use more template. Sample concentration may be too low.
- Template DNA may be damaged. Use carefully purified template.
- Lengthen extension time.
- Increase cycle number.
- Denaturation temperature may be too low. Optimal denaturation temperature for most templates is 98°C or higher.
Find a Golden Butterfly for a Chance to Win!
Help us celebrate our 50th anniversary! We have hidden 1,000 golden butterflies and are waiting for you to find them. They can be anywhere that you find NEB! Beginning April 15th, be sure to visit our website and tables at tradeshows and events you are attending. Visit our social media channels frequently for tips on where we have hidden the butterflies – and once you find one, either click or scan the code to be eligible for a 50th anniversary prize pack, as well as a grand prize trip to NEB headquarters in Ipswich, MA!.
