|
PCR additives Betaine, DMSO and formamide reduces the Tm and the complex secondary structure thus the duplex stability. Tetramethyl ammonium chloride
(TMAC) actually increases the specificity of hybridization and increases the Tm. The use of TMAC is recommended in PCR conditions using degenerate primers.
These PCR additives and enhancing agents have been used to increase the yield, specificity and
consistency of PCR reactions. These additives may have beneficial effects on some amplification and
it is impossible to predict which agents will be useful in a particular context and therefore they must be empirically tested for each combination of template and primers.
Betaine reduces Tm facilitating GC rich region
amplification. Reduces duplex stability.Use 3.5M to 0.1M betaine. Be sure to use Betaine or Betaine (mono)hydrate and not
Betaine HCl.
|
