Owing to a uniquely balanced combination of KCl and (NH 4) 2SO 4, the PCR buffer provides stringent primer annealing conditions over a wider range of annealing temperatures and Mg 2+ concentrations, compared with conventional PCR buffers. The high yields of specific PCR products obtained ensure reliable Pyrosequencing results.Дuring the annealing step of every PCR cycle, the buffer composition ensures a high ratio of specific-to-nonspecific primer binding (see figure ' Increased specificity of primer annealing'). The special formulation also prevents accumulation of excess biotinylated primer and minimizes generation of artifacts that can interfere with the Pyrosequencing reaction. The balanced combination of salts in the PyroMark PCR Kit's unique master mix ensures specifically targeted primer binding and prevents mispriming.

This less complex DNA is a challenging PCR template - the PCR products obtained are likely to have low yields, and nonspecific products might also be generated due to increased probability for mispriming. Bisulfite treatment of genomic DNA converts nonmethylated cytosines to uracils andВ produces DNA that mainly consists of three bases. The convenient master mix format enables specific amplification of bisulfite converted DNA or genomic DNA from various starting materials, using only one protocol. The PyroMark PCR Kit enables PCR amplification of template DNA optimized for Pyrosequencing analysis.

Pcr cheek cell lab master mix