Designing Primers - A Checklist

Biochemistry, Molecular Biology, and Cell Biology Protocols >> Designing Primers - A Checklist

Are annealing temperatures of each pair of primers for a corresponding PCR product similar?

Does the primer have
   an annealing segment?
   a tag segment?
   a restriction enzyme site?
   extra bases to help cut the restriction site?

Is the primer in the correct orientation (if necessary, is it in reverse complement? If necessary, are the extra bases [d] in reverse complement?)

Is the annealing temperature at least 50 degrees Celsius?

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