There are many points to consider when you carry out LA PCR; for example, the pH of

reaction buffer, salt concentration, reaction conditions in cycling, primer design and so on.

Before beginning PCR, preliminary experiments to optimize your reaction conditions are

essential. PCR success heavily depends upon your individual template-primer combinations.

Conditions for optimization may include cycling parameters and concentrations of essential

reaction components; Mg, dNTP, primers, and DNA polymerase.

 

circle07_blue.gif  Template DNA
circle07_blue.gif  Protocol for the preparation of human genomic DNA
circle07_blue.gif  Protocol for the preparation of E. coli genomic DNA
circle07_blue.gif  Primer preparation
circle07_blue.gif  Primer concentration
circle07_blue.gif  Magenesium concentration
circle07_blue.gif  Enzyme amount
circle07_blue.gif  Cycle numbers
circle07_blue.gif  Denaturation conditions
circle07_blue.gif  Conditions for annealing and extension
circle07_blue.gif  Hot Start method