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Using the tool Gene2Oligo I have a set of DNA oligonucleotides to synthesize a gene using ligase chain reaction (LCR). The average melting temperature is 72 degrees Celsius. I am going to use a the thermostable Taq ligase enzyme for the LCR reaction but I am unsure of the annealing temperature to use. Should I use the standard recommendation as in PCR of 5 degrees less than the melting temperature or should I use another tool? Taq ligase has a cutoff in activity at 65C so that is the highest I can use.

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