PRIMER_MAX_POLY_X=4 # Max mononucleotide repeats (e.g., AAAA) PRIMER_GC_CLAMP=1 # Require G or C in last 5 bases of 3' end PRIMER_MAX_SELF_ANY=4.0 PRIMER_MAX_SELF_END=3.0 PRIMER_NUM_RETURN=5
But Primer3 is not a "point-and-click" black box. It is a . To unlock its full potential, you must understand its input format. primer3 input
SEQUENCE_INTERNAL_OLIGO_MIN_SIZE=18 SEQUENCE_INTERNAL_OLIGO_OPT_TM=55.0 Save this as my_primers.txt : PRIMER_MAX_POLY_X=4 # Max mononucleotide repeats (e