Primer3 Output ------------------------------------------------------------------------ WARNING: Numbers in input sequence were deleted. No mispriming library specified Using 1-based sequence positions OLIGO start len tm gc% any 3' seq LEFT PRIMER 60 20 60.02 50.00 5.00 2.00 acgtcttgctcagcctcaat RIGHT PRIMER 243 20 60.32 55.00 5.00 1.00 gttacctcttgcggcagtgt SEQUENCE SIZE: 460 INCLUDED REGION SIZE: 460 PRODUCT SIZE: 184, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 3.00 1 caggatgtctgtgagtgagagtgcagtgtttgcctacgagtcctctgtgcacagcaccaa > 61 cgtcttgctcagcctcaatgaccagcggaagaaggatgtgctgtgcgatgtcactgtcct >>>>>>>>>>>>>>>>>>> 121 ggtggagggccagcagttccgagcccaccgctcggtgctggctgcatgcagcagctactt 181 ccactcgcgaatcgtaggccaaactgatgccgagctcaccatcacactgccgcaagaggt <<<<<<<<<<<<<<<<< 241 aacagttaaaggatttgaacctttaattcagtttgcctacactgccaaactcattttaag <<< 301 taaagacaatgttgacgaagtctgcaagtgtgtggagtttctaagcgtacacaatatcga 361 ggaatcctgctttcagtttctcaagtttaagtttttggactccacctcagagcagcaggg 421 atgtgcaagaaaaaagtgcttctcctcacactgtcagaaa KEYS (in order of precedence): >>>>>> left primer <<<<<< right primer ADDITIONAL OLIGOS start len tm gc% any 3' seq 1 LEFT PRIMER 60 20 60.02 50.00 5.00 2.00 acgtcttgctcagcctcaat RIGHT PRIMER 287 20 60.45 50.00 5.00 3.00 tggcagtgtaggcaaactga PRODUCT SIZE: 228, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 3.00 2 LEFT PRIMER 60 20 60.02 50.00 5.00 2.00 acgtcttgctcagcctcaat RIGHT PRIMER 245 20 60.45 55.00 5.00 1.00 ctgttacctcttgcggcagt PRODUCT SIZE: 186, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 3.00 3 LEFT PRIMER 191 20 60.10 50.00 4.00 2.00 atcgtaggccaaactgatgc RIGHT PRIMER 413 20 60.39 55.00 7.00 3.00 gctctgaggtggagtccaaa PRODUCT SIZE: 223, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 0.00 4 LEFT PRIMER 20 20 59.51 50.00 4.00 0.00 agtgcagtgtttgcctacga RIGHT PRIMER 193 20 59.98 55.00 6.00 2.00 gattcgcgagtggaagtagc PRODUCT SIZE: 174, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00 Statistics con too in in no tm tm high high high sid many tar excl bad GC too too any 3' poly end ered Ns get reg GC% clamp low high compl compl X stab ok Left 2805 0 0 0 3 0 613 1573 2 1 0 36 577 Right 2739 0 0 0 3 0 900 1104 1 3 19 31 678 Pair Stats: considered 55, unacceptable product size 46, ok 9 primer3 release 1.0 (primer3_www_results.cgi v 0.4)