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Please use ACGTN symbols for the primer sequences, IUPAC codes (BDHRYKMSWV) for degenerate bases and “I” for Inosine.
If there are any incomplementary overlaps at 5’ ends of primers (e.g. with a restriction site), please enter only the portion of the primer sequence which is complementary to the sequenced template.
I really appreciate your extremely helpful approach, I will praise you to all colleagues to heaven.
Karolina
Biology Centre CAS, Ceske Budejovice