Concerning the Primer

• It is the aberrant binding of the primer. It carries the primer sequence and becomes a target for the next amplifications.

  Misprimes
• These are artifacts in PCR, they are PCR products that double in size of primers, and they are primers that bind with each other.

  Primer dimers
• Primers bind with each other because Taq polymerase has some activity at room temperature(22-25 degrees celcius) and at the same time, primers can bind sequences other than their exact complements on the target (low stringency).
• Solution 1: Good Preparation consists of?

  Good primer design

  Optimal amplification conditions
• Solution 2: Hot-Start PCR types.

  Type 1

  Type 2 (Wax Barrier)

  Type 3
• Hot-Start PCR type: Reaction mixes are prepped on ice and placed in a prewarmed thermal cycler.

  Type 1
• Hot-Start PCR type: Wax separates the enzyme and template from the primers.

  Type 2 (Wax barrier)
• HOT-Start PCR type: Supplied in inactive form.

  Type 3