Effect Of Temperature & Salt Concentration; Summary Of Conditions Affecting Hybridisation - Thermo Scientific Shake n Stack 6240 Operating And Maintenance Manual

Effect of Temperature & Salt Concentration
HIGH
LOW
Temperature/Monovalentation concentration
- - - - - - Temperature
Monovalentation

Summary of Conditions Affecting Hybridisation

Temperature
The Hybridisation temperature is one of the most critical factors because it affects the
rate of hydrogen bond formation between DNA strands; the hydrogen bonds in effect
overcome the repulsive forces between the negatively charged strands. If the
temperature is increased, then the stringency of Hybridisation is increased.
Ionic Strength
Increasing the monovalent cation concentration enables the DNA strands to come
together more readily, thus reducing the stringency of Hybridisation. EDTA is sometimes
added to chelate magnesium ions, which can have an unpredictable effect on
Hybridisation reactions.
© Thermo Scientific, May 2003. Issue 7
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