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The page below is a sample from the LabCE course Fundamentals of Molecular Diagnostics. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Factors Affecting Hybridization

Because hybridization involves nucleotide bases and the separation and joining or reannealing of strands, several environmental factors can influence this process:

  • Temperature: If the temperature is too high, the strands melt. If it is too low, they might be forced together.
  • The pH: A pH that is too alkaline will cause the strands to separate; too acidic and they are forced together.
  • The guanine to cytosine ratio (G:C ratio): Since this bond is stronger than the other nucleotide bonds, if the G:C ratio in the desired target strand is high, the separation process may take longer.