Objective To discuss the SOE-PCR technique in long piece or multiple pieces gene mutations operation.Methods After the primary PCR reaction,besides putting the flanking primers of recommended concentration,we changed the traditional way into putting 100-time diluted internal primers which used to amplify the primary PCR templates.Through such operations,we can achieve the most satisfied templates concentration of SOE-PCR dynamically.Results Compare with the traditional SOE-PCR,two pieces combination products with the new way is more stable and specific while the specific abundance ratio keeps well.At the same time we achieve the three pieces combination with a total length of 4.2 kb in the same reaction while the specific amplification abundance also satisfied.Conclusion With the new method,we can overlap pieces combination of SOE-PCR in the same reaction tube with the specificity and abundance keeps well while not like the traditional way of combine two pieces together.When in multiple chimerical genes operation,this kind of method can reduce PCR reaction cycles,thus may be meaningful for chimerical genes operations.
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