Causes

Trouble-shooting measures

  Enzyme concentration is too high.

Reduce the amount in decrements of 0.5 U.

  Denaturation time is too short.

Elongate the denaturation in increments of

5 seconds.

  Denaturation temperature is low.

Raise the temperature in increments of 0.5.

  Concentration of dNTP is low.

Increase the concentration in increments of

50 .

  Extension time is too long.

Shorten the extension time in decrements of

1 minute.

  Cycle number is too high.

Reduce the number of cycles in decrements of

2 cycles.

  Template concentration is too high.

Reduce the amount in decrements of 20% of

the previous one.