Target amplification methods enzymes needed

Since rolling circle amplification takes place at a constant temperature, there is no need for the target amplification process to take place in a thermal cycler, which is required to regulate the temperature for different parts of the reaction. The type of DNA polymerase to be used in RCA is not limited to thermostable enzymes, like the PCR-based diagnostics. On the other hand, the RCA method requires the environment to be free of contaminations as the RCA arrays are highly sensitive. Wiltshire [22]... 

Amplification assays can produce both qualitative answers (yes/no, more or less abundant than reference materials) and quantitative answers (the original concentration of target sequence in the sample). In order to use the amplification process for quantitative assessments, many variables need to be carefully controlled. Variations in extraction efficiency, presence of enzyme inhibitors, lot-to-lot variation in enzyme and reagent performance, and day-to-day variation in reaction and detection conditions need to be addressed in methods that attempt to yield a quantitative result. 

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