P3_COMMENT=Test SEQUENCE_OVERHANG_LEFT and SEQUENCE_OVERHANG_RIGHT
PRIMER_TASK=check_primers
PRIMER_PICK_ANYWAY=1
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_MAX_NS_ACCEPTED=100
PRIMER_MAX_SIZE=35
PRIMER_MIN_SIZE=14
PRIMER_PRODUCT_SIZE_RANGE=60-1000
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=1
SEQUENCE_ID=Overhang01
SEQUENCE_TEMPLATE=GACATACGATACTTTGCTTTCAATCATGTGGGTGAAACGACTAAACCTTACGACTACTTAAAGTGGACTGGTAGTCTTTTTAAAGGTAC
SEQUENCE_PRIMER=TGCTTTCAATCATGTGGGTGA
SEQUENCE_OVERHANG_LEFT=tcacccac
SEQUENCE_PRIMER_REVCOMP=GACTACCAGTCCACTTTAAG
SEQUENCE_OVERHANG_RIGHT=TAAGGATTT
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=10.192729
PRIMER_LEFT_0_PENALTY=2.874014
PRIMER_RIGHT_0_PENALTY=7.318716
PRIMER_LEFT_0_PROBLEMS= Hairpin stability too high;
PRIMER_RIGHT_0_PROBLEMS= Temperature too low;
PRIMER_LEFT_0_SEQUENCE=TCACCCACTGCTTTCAATCATGTGGGTGA
PRIMER_RIGHT_0_SEQUENCE=TAAGGATTTGACTACCAGTCCACTTTAAG
PRIMER_LEFT_0=14,21
PRIMER_RIGHT_0=75,20
PRIMER_LEFT_0_TM=58.126
PRIMER_RIGHT_0_TM=52.681
PRIMER_LEFT_0_GC_PERCENT=42.857
PRIMER_RIGHT_0_GC_PERCENT=45.000
PRIMER_LEFT_0_SELF_ANY_TH=38.38
PRIMER_RIGHT_0_SELF_ANY_TH=3.97
PRIMER_LEFT_0_SELF_ANY_STUCT=Tm: 38.4&deg;C  dG: -11765 cal/mol  dH: -123600 cal/mol  dS: -361 cal/mol*K\n5' TCACCCACTGCTTTCAATCATGTGGGTGA 3'\n   ||||||||             ||||||||\n3' AGTGGGTGTACTAACTTTCGTCACCCACT 5'\n
PRIMER_RIGHT_0_SELF_ANY_STUCT=Tm: 4.0&deg;C  dG: -3087 cal/mol  dH: -68200 cal/mol  dS: -210 cal/mol*K\n5' TAAGGATTTGACTACCAGTCCACTTTAAG 3'\n            ||||   ||||\n3' GAATTTCACCTGACCATCAGTTTAGGAAT 5'\n
PRIMER_LEFT_0_SELF_END_TH=38.38
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_SELF_END_STUCT=Tm: 38.4&deg;C  dG: -11765 cal/mol  dH: -123600 cal/mol  dS: -361 cal/mol*K\n5' TCACCCACTGCTTTCAATCATGTGGGTGA 3'\n   ||||||||             ||||||||\n3' AGTGGGTGTACTAACTTTCGTCACCCACT 5'\n
PRIMER_LEFT_0_HAIRPIN_TH=65.45
PRIMER_RIGHT_0_HAIRPIN_TH=44.44
PRIMER_LEFT_0_HAIRPIN_STUCT=Tm: 65.4&deg;C  dG: -4873 cal/mol  dH: -58000 cal/mol  dS: -171 cal/mol*K\n5' TCACCCACTGCTTTU+2510\n    |||||||      C\n3' AGTGGGTGTACTAAU+2518\n
PRIMER_RIGHT_0_HAIRPIN_STUCT=Tm: 44.4&deg;C  dG: -595 cal/mol  dH: -25400 cal/mol  dS: -80 cal/mol*K\n5' TAAGGATTTGACTAU+2510\n            |||| C\n3' GAATTTCACCTGACU+2518\n
PRIMER_LEFT_0_END_STABILITY=4.0200
PRIMER_RIGHT_0_END_STABILITY=1.8500
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=79
PRIMER_PAIR_0_PRODUCT_TM=76.0
=
SEQUENCE_ID=Overhang02
PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=1
SEQUENCE_TEMPLATE=TCACCCACTGCTTTCAATCATGTGGGTGAAACGACTAAACCTTACGACTACTTAAAGTGGACTGGTAGTCAAATCCTTA
SEQUENCE_PRIMER=TCACCCACTGCTTTCAATCATGTGGGTGA
SEQUENCE_PRIMER_REVCOMP=TAAGGATTTGACTACCAGTCCACTTTAAG
P3_COMMENT=same oligos as Overhang01 but without using SEQUENCE_OVERHANG_LEFT/RIGHT
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=27.157692
PRIMER_LEFT_0_PENALTY=17.215227
PRIMER_RIGHT_0_PENALTY=9.942466
PRIMER_LEFT_0_PROBLEMS= Temperature too high; Hairpin stability too high;
PRIMER_LEFT_0_SEQUENCE=TCACCCACTGCTTTCAATCATGTGGGTGA
PRIMER_RIGHT_0_SEQUENCE=TAAGGATTTGACTACCAGTCCACTTTAAG
PRIMER_LEFT_0=0,29
PRIMER_RIGHT_0=78,29
PRIMER_LEFT_0_TM=68.215
PRIMER_RIGHT_0_TM=60.942
PRIMER_LEFT_0_GC_PERCENT=48.276
PRIMER_RIGHT_0_GC_PERCENT=37.931
PRIMER_LEFT_0_SELF_ANY_TH=38.38
PRIMER_RIGHT_0_SELF_ANY_TH=3.97
PRIMER_LEFT_0_SELF_ANY_STUCT=Tm: 38.4&deg;C  dG: -11765 cal/mol  dH: -123600 cal/mol  dS: -361 cal/mol*K\n5' TCACCCACTGCTTTCAATCATGTGGGTGA 3'\n   ||||||||             ||||||||\n3' AGTGGGTGTACTAACTTTCGTCACCCACT 5'\n
PRIMER_RIGHT_0_SELF_ANY_STUCT=Tm: 4.0&deg;C  dG: -3087 cal/mol  dH: -68200 cal/mol  dS: -210 cal/mol*K\n5' TAAGGATTTGACTACCAGTCCACTTTAAG 3'\n            ||||   ||||\n3' GAATTTCACCTGACCATCAGTTTAGGAAT 5'\n
PRIMER_LEFT_0_SELF_END_TH=38.38
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_SELF_END_STUCT=Tm: 38.4&deg;C  dG: -11765 cal/mol  dH: -123600 cal/mol  dS: -361 cal/mol*K\n5' TCACCCACTGCTTTCAATCATGTGGGTGA 3'\n   ||||||||             ||||||||\n3' AGTGGGTGTACTAACTTTCGTCACCCACT 5'\n
PRIMER_LEFT_0_HAIRPIN_TH=65.45
PRIMER_RIGHT_0_HAIRPIN_TH=44.44
PRIMER_LEFT_0_HAIRPIN_STUCT=Tm: 65.4&deg;C  dG: -4873 cal/mol  dH: -58000 cal/mol  dS: -171 cal/mol*K\n5' TCACCCACTGCTTTU+2510\n    |||||||      C\n3' AGTGGGTGTACTAAU+2518\n
PRIMER_RIGHT_0_HAIRPIN_STUCT=Tm: 44.4&deg;C  dG: -595 cal/mol  dH: -25400 cal/mol  dS: -80 cal/mol*K\n5' TAAGGATTTGACTAU+2510\n            |||| C\n3' GAATTTCACCTGACU+2518\n
PRIMER_LEFT_0_END_STABILITY=4.0200
PRIMER_RIGHT_0_END_STABILITY=1.8500
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=79
PRIMER_PAIR_0_PRODUCT_TM=78.0
=
SEQUENCE_ID=Overhang03
PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=0
SEQUENCE_TEMPLATE=GACATACGATACTTTGCTTTCAATCATGTGGGTGAAACGACTAAACCTTACGACTACTTAAAGTGGACTGGTAGTCTTTTTAAAGGTAC
SEQUENCE_PRIMER=TGCTTTCAATCATGTGGGTGA
SEQUENCE_OVERHANG_LEFT=TCACCCAC
SEQUENCE_PRIMER_REVCOMP=GACTACCAGTCCACTTTAAG
SEQUENCE_OVERHANG_RIGHT=TAAGGATTT
P3_COMMENT=same as Overhang01 but with PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=0
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=10.192729
PRIMER_LEFT_0_PENALTY=2.874014
PRIMER_RIGHT_0_PENALTY=7.318716
PRIMER_LEFT_0_PROBLEMS= Hairpin stability too high;
PRIMER_RIGHT_0_PROBLEMS= Temperature too low;
PRIMER_LEFT_0_SEQUENCE=TCACCCACTGCTTTCAATCATGTGGGTGA
PRIMER_RIGHT_0_SEQUENCE=TAAGGATTTGACTACCAGTCCACTTTAAG
PRIMER_LEFT_0=14,21
PRIMER_RIGHT_0=75,20
PRIMER_LEFT_0_TM=58.126
PRIMER_RIGHT_0_TM=52.681
PRIMER_LEFT_0_GC_PERCENT=42.857
PRIMER_RIGHT_0_GC_PERCENT=45.000
PRIMER_LEFT_0_SELF_ANY_TH=38.38
PRIMER_RIGHT_0_SELF_ANY_TH=3.97
PRIMER_LEFT_0_SELF_END_TH=38.38
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=65.45
PRIMER_RIGHT_0_HAIRPIN_TH=44.44
PRIMER_LEFT_0_END_STABILITY=4.0200
PRIMER_RIGHT_0_END_STABILITY=1.8500
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=79
PRIMER_PAIR_0_PRODUCT_TM=76.0
=
SEQUENCE_ID=Overhang04
PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=0
SEQUENCE_TEMPLATE=TCACCCACTGCTTTCAATCATGTGGGTGAAACGACTAAACCTTACGACTACTTAAAGTGGACTGGTAGTCAAATCCTTA
SEQUENCE_PRIMER=TCACCCACTGCTTTCAATCATGTGGGTGA
SEQUENCE_PRIMER_REVCOMP=TAAGGATTTGACTACCAGTCCACTTTAAG
P3_COMMENT=same as Overhang02 but with PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=0
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=27.157692
PRIMER_LEFT_0_PENALTY=17.215227
PRIMER_RIGHT_0_PENALTY=9.942466
PRIMER_LEFT_0_PROBLEMS= Temperature too high; Hairpin stability too high;
PRIMER_LEFT_0_SEQUENCE=TCACCCACTGCTTTCAATCATGTGGGTGA
PRIMER_RIGHT_0_SEQUENCE=TAAGGATTTGACTACCAGTCCACTTTAAG
PRIMER_LEFT_0=0,29
PRIMER_RIGHT_0=78,29
PRIMER_LEFT_0_TM=68.215
PRIMER_RIGHT_0_TM=60.942
PRIMER_LEFT_0_GC_PERCENT=48.276
PRIMER_RIGHT_0_GC_PERCENT=37.931
PRIMER_LEFT_0_SELF_ANY_TH=38.38
PRIMER_RIGHT_0_SELF_ANY_TH=3.97
PRIMER_LEFT_0_SELF_END_TH=38.38
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=65.45
PRIMER_RIGHT_0_HAIRPIN_TH=44.44
PRIMER_LEFT_0_END_STABILITY=4.0200
PRIMER_RIGHT_0_END_STABILITY=1.8500
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=79
PRIMER_PAIR_0_PRODUCT_TM=78.0
=
SEQUENCE_ID=Overhang05
PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=1
SEQUENCE_TEMPLATE=GGACGTCAGCGGGTGACGTCCCCCCTTGTAAGATACGCGGGTGACGACCCCCATCAACAATCAATCGTTTATAATCCTTATTTCTGACAGGCTGAAGGACTCAGGACTCCCCATAAGAAATTGTTCGGGGGACGTCACCACTCAGTGAGA
SEQUENCE_PRIMER=GCGGGTGACGTCCCCC
SEQUENCE_OVERHANG_LEFT=GGACGTCA
SEQUENCE_PRIMER_REVCOMP=GAACAATTTCTTATGGGG
SEQUENCE_OVERHANG_RIGHT=GGTGACGTCCCCC
P3_COMMENT=overhangs and full oligos have hairpins and complementary regions
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=20.519295
PRIMER_LEFT_0_PENALTY=6.314471
PRIMER_RIGHT_0_PENALTY=14.204824
PRIMER_LEFT_0_PROBLEMS= GC content too high; Similarity to self too high; Hairpin stability too high;
PRIMER_RIGHT_0_PROBLEMS= Temperature too low;
PRIMER_LEFT_0_SEQUENCE=GGACGTCAGCGGGTGACGTCCCCC
PRIMER_RIGHT_0_SEQUENCE=GGTGACGTCCCCCGAACAATTTCTTATGGGG
PRIMER_LEFT_0=8,16
PRIMER_RIGHT_0=125,18
PRIMER_LEFT_0_TM=62.314
PRIMER_RIGHT_0_TM=47.795
PRIMER_LEFT_0_GC_PERCENT=81.250
PRIMER_RIGHT_0_GC_PERCENT=38.889
PRIMER_LEFT_0_SELF_ANY_TH=55.63
PRIMER_RIGHT_0_SELF_ANY_TH=14.12
PRIMER_LEFT_0_SELF_ANY_STUCT=Tm: 55.6&deg;C  dG: -19920 cal/mol  dH: -153600 cal/mol  dS: -431 cal/mol*K\n   5' GGACGTCAGCGGGTGACGTCCCCC 3'\n      |||||||| | | ||||||||\n3' CCCCCTGCAGTGGGCGACTGCAGG 5'\n
PRIMER_RIGHT_0_SELF_ANY_STUCT=Tm: 14.1&deg;C  dG: -6548 cal/mol  dH: -58600 cal/mol  dS: -168 cal/mol*K\n                   5' GGTGACGTCCCCCGAACAATTTCTTATGGGG 3'\n                         ||||||\n3' GGGGTATTCTTTAACAAGCCCCCTGCAGTGG 5'\n
PRIMER_LEFT_0_SELF_END_TH=45.91
PRIMER_RIGHT_0_SELF_END_TH=6.00
PRIMER_LEFT_0_SELF_END_STUCT=Tm: 45.9&deg;C  dG: -15142 cal/mol  dH: -140600 cal/mol  dS: -405 cal/mol*K\n   5' GGACGTCAGCGGGTGACGTCCCCC 3'\n      |||||||| | | ||||||   ||\n3' CCCCCTGCAGTGGGCGACTGCA---GG 5'\n
PRIMER_RIGHT_0_SELF_END_STUCT=Tm: 6.0&deg;C  dG: -4087 cal/mol  dH: -64200 cal/mol  dS: -194 cal/mol*K\n5' GGTGACGTCCCCCGAACAATTTCTTATGGGG 3'\n            ||||              ||||\n         3' GGGGTATTCTTTAACAAGCCCCCTGCAGTGG 5'\n
PRIMER_LEFT_0_HAIRPIN_TH=74.07
PRIMER_RIGHT_0_HAIRPIN_TH=43.63
PRIMER_LEFT_0_HAIRPIN_STUCT=Tm: 74.1&deg;C  dG: -4121 cal/mol  dH: -38600 cal/mol  dS: -111 cal/mol*K\n   5' GGACGTCAGCU+2510\n        ||||||  G\n3' CCCCCTGCAGTGGU+2518\n
PRIMER_RIGHT_0_HAIRPIN_STUCT=Tm: 43.6&deg;C  dG: -950 cal/mol  dH: -45400 cal/mol  dS: -143 cal/mol*K\n5' GGTGACGTCCCCCGAACAU+2510\n            |||||||  U+2502\n     3' GGGGTATTCTTTAU+2518\n
PRIMER_LEFT_0_END_STABILITY=5.4000
PRIMER_RIGHT_0_END_STABILITY=4.9600
PRIMER_PAIR_0_COMPL_ANY_TH=34.86
PRIMER_PAIR_0_COMPL_ANY_STUCT=Tm: 34.9&deg;C  dG: -10657 cal/mol  dH: -80500 cal/mol  dS: -225 cal/mol*K\n        5' GGACGTCAGCGGGTGACGTCCCCC 3'\n                    |||| ||||||\n3' GGGGTATTCTTTAACAAGCCCCCTGCAGTGG 5'\n
PRIMER_PAIR_0_COMPL_END_TH=44.57
PRIMER_PAIR_0_COMPL_END_STUCT=Tm: 44.6&deg;C  dG: -13298 cal/mol  dH: -87400 cal/mol  dS: -239 cal/mol*K\n5' CCCCATAAGAAATTGTTCGGGGGACGTCACC 3'\n                    |||| |||||| ||\n        3' CCTGCAGTCGCCCACTGCAGGGGG 5'\n
PRIMER_PAIR_0_PRODUCT_SIZE=139
PRIMER_PAIR_0_PRODUCT_TM=83.5
=
SEQUENCE_ID=Overhang06
PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=1
SEQUENCE_TEMPLATE=GGACGTCAGCGGGTGACGTCCCCCCTTGTAAGATACGCGGGTGACGACCCCCATCAACAATCAATCGTTTATAATCCTTATTTCTGACAGGCTGAAGGACTCAGGACTCCCCATAAGAAATTGTTCGGGGGACGTCACCACTCAGTGAGA
SEQUENCE_PRIMER=GGACGTCAGCGGGTGACGTCCCCC
SEQUENCE_PRIMER_REVCOMP=GGTGACGTCCCCCGAACAATTTCTTATGGGG
P3_COMMENT=same oligos as Overhang05 but without using SEQUENCE_OVERHANG_LEFT/RIGHT
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=37.906285
PRIMER_LEFT_0_PENALTY=16.725787
PRIMER_RIGHT_0_PENALTY=21.180499
PRIMER_LEFT_0_PROBLEMS= Temperature too high; Similarity to self too high; Hairpin stability too high;
PRIMER_RIGHT_0_PROBLEMS= Temperature too high;
PRIMER_LEFT_0_SEQUENCE=GGACGTCAGCGGGTGACGTCCCCC
PRIMER_RIGHT_0_SEQUENCE=GGTGACGTCCCCCGAACAATTTCTTATGGGG
PRIMER_LEFT_0=0,24
PRIMER_RIGHT_0=138,31
PRIMER_LEFT_0_TM=72.726
PRIMER_RIGHT_0_TM=70.180
PRIMER_LEFT_0_GC_PERCENT=75.000
PRIMER_RIGHT_0_GC_PERCENT=54.839
PRIMER_LEFT_0_SELF_ANY_TH=55.63
PRIMER_RIGHT_0_SELF_ANY_TH=14.12
PRIMER_LEFT_0_SELF_ANY_STUCT=Tm: 55.6&deg;C  dG: -19920 cal/mol  dH: -153600 cal/mol  dS: -431 cal/mol*K\n   5' GGACGTCAGCGGGTGACGTCCCCC 3'\n      |||||||| | | ||||||||\n3' CCCCCTGCAGTGGGCGACTGCAGG 5'\n
PRIMER_RIGHT_0_SELF_ANY_STUCT=Tm: 14.1&deg;C  dG: -6548 cal/mol  dH: -58600 cal/mol  dS: -168 cal/mol*K\n                   5' GGTGACGTCCCCCGAACAATTTCTTATGGGG 3'\n                         ||||||\n3' GGGGTATTCTTTAACAAGCCCCCTGCAGTGG 5'\n
PRIMER_LEFT_0_SELF_END_TH=45.91
PRIMER_RIGHT_0_SELF_END_TH=6.00
PRIMER_LEFT_0_SELF_END_STUCT=Tm: 45.9&deg;C  dG: -15142 cal/mol  dH: -140600 cal/mol  dS: -405 cal/mol*K\n   5' GGACGTCAGCGGGTGACGTCCCCC 3'\n      |||||||| | | ||||||   ||\n3' CCCCCTGCAGTGGGCGACTGCA---GG 5'\n
PRIMER_RIGHT_0_SELF_END_STUCT=Tm: 6.0&deg;C  dG: -4087 cal/mol  dH: -64200 cal/mol  dS: -194 cal/mol*K\n5' GGTGACGTCCCCCGAACAATTTCTTATGGGG 3'\n            ||||              ||||\n         3' GGGGTATTCTTTAACAAGCCCCCTGCAGTGG 5'\n
PRIMER_LEFT_0_HAIRPIN_TH=74.07
PRIMER_RIGHT_0_HAIRPIN_TH=43.63
PRIMER_LEFT_0_HAIRPIN_STUCT=Tm: 74.1&deg;C  dG: -4121 cal/mol  dH: -38600 cal/mol  dS: -111 cal/mol*K\n   5' GGACGTCAGCU+2510\n        ||||||  G\n3' CCCCCTGCAGTGGU+2518\n
PRIMER_RIGHT_0_HAIRPIN_STUCT=Tm: 43.6&deg;C  dG: -950 cal/mol  dH: -45400 cal/mol  dS: -143 cal/mol*K\n5' GGTGACGTCCCCCGAACAU+2510\n            |||||||  U+2502\n     3' GGGGTATTCTTTAU+2518\n
PRIMER_LEFT_0_END_STABILITY=5.4000
PRIMER_RIGHT_0_END_STABILITY=4.9600
PRIMER_PAIR_0_COMPL_ANY_TH=34.86
PRIMER_PAIR_0_COMPL_ANY_STUCT=Tm: 34.9&deg;C  dG: -10657 cal/mol  dH: -80500 cal/mol  dS: -225 cal/mol*K\n        5' GGACGTCAGCGGGTGACGTCCCCC 3'\n                    |||| ||||||\n3' GGGGTATTCTTTAACAAGCCCCCTGCAGTGG 5'\n
PRIMER_PAIR_0_COMPL_END_TH=44.57
PRIMER_PAIR_0_COMPL_END_STUCT=Tm: 44.6&deg;C  dG: -13298 cal/mol  dH: -87400 cal/mol  dS: -239 cal/mol*K\n5' CCCCATAAGAAATTGTTCGGGGGACGTCACC 3'\n                    |||| |||||| ||\n        3' CCTGCAGTCGCCCACTGCAGGGGG 5'\n
PRIMER_PAIR_0_PRODUCT_SIZE=139
PRIMER_PAIR_0_PRODUCT_TM=85.7
=
SEQUENCE_ID=Overhang07
PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=0
SEQUENCE_TEMPLATE=GGACGTCAGCGGGTGACGTCCCCCCTTGTAAGATACGCGGGTGACGACCCCCATCAACAATCAATCGTTTATAATCCTTATTTCTGACAGGCTGAAGGACTCAGGACTCCCCATAAGAAATTGTTCGGGGGACGTCACCACTCAGTGAGA
SEQUENCE_PRIMER=GCGGGTGACGTCCCCC
SEQUENCE_OVERHANG_LEFT=GGACGTCA
SEQUENCE_PRIMER_REVCOMP=GAACAATTTCTTATGGGG
SEQUENCE_OVERHANG_RIGHT=GGTGACGTCCCCC
P3_COMMENT=same as Overhang05 but with PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=0
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=20.519295
PRIMER_LEFT_0_PENALTY=6.314471
PRIMER_RIGHT_0_PENALTY=14.204824
PRIMER_LEFT_0_PROBLEMS= GC content too high; Similarity to self too high; Hairpin stability too high;
PRIMER_RIGHT_0_PROBLEMS= Temperature too low;
PRIMER_LEFT_0_SEQUENCE=GGACGTCAGCGGGTGACGTCCCCC
PRIMER_RIGHT_0_SEQUENCE=GGTGACGTCCCCCGAACAATTTCTTATGGGG
PRIMER_LEFT_0=8,16
PRIMER_RIGHT_0=125,18
PRIMER_LEFT_0_TM=62.314
PRIMER_RIGHT_0_TM=47.795
PRIMER_LEFT_0_GC_PERCENT=81.250
PRIMER_RIGHT_0_GC_PERCENT=38.889
PRIMER_LEFT_0_SELF_ANY_TH=55.63
PRIMER_RIGHT_0_SELF_ANY_TH=14.12
PRIMER_LEFT_0_SELF_END_TH=45.91
PRIMER_RIGHT_0_SELF_END_TH=6.00
PRIMER_LEFT_0_HAIRPIN_TH=74.07
PRIMER_RIGHT_0_HAIRPIN_TH=43.63
PRIMER_LEFT_0_END_STABILITY=5.4000
PRIMER_RIGHT_0_END_STABILITY=4.9600
PRIMER_PAIR_0_COMPL_ANY_TH=34.86
PRIMER_PAIR_0_COMPL_END_TH=44.57
PRIMER_PAIR_0_PRODUCT_SIZE=139
PRIMER_PAIR_0_PRODUCT_TM=83.5
=
SEQUENCE_ID=Overhang08
PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=0
SEQUENCE_TEMPLATE=GGACGTCAGCGGGTGACGTCCCCCCTTGTAAGATACGCGGGTGACGACCCCCATCAACAATCAATCGTTTATAATCCTTATTTCTGACAGGCTGAAGGACTCAGGACTCCCCATAAGAAATTGTTCGGGGGACGTCACCACTCAGTGAGA
SEQUENCE_PRIMER=GGACGTCAGCGGGTGACGTCCCCC
SEQUENCE_PRIMER_REVCOMP=GGTGACGTCCCCCGAACAATTTCTTATGGGG
P3_COMMENT=same oligos as Overhang07 but without using SEQUENCE_OVERHANG_LEFT/RIGHT
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=37.906285
PRIMER_LEFT_0_PENALTY=16.725787
PRIMER_RIGHT_0_PENALTY=21.180499
PRIMER_LEFT_0_PROBLEMS= Temperature too high; Similarity to self too high; Hairpin stability too high;
PRIMER_RIGHT_0_PROBLEMS= Temperature too high;
PRIMER_LEFT_0_SEQUENCE=GGACGTCAGCGGGTGACGTCCCCC
PRIMER_RIGHT_0_SEQUENCE=GGTGACGTCCCCCGAACAATTTCTTATGGGG
PRIMER_LEFT_0=0,24
PRIMER_RIGHT_0=138,31
PRIMER_LEFT_0_TM=72.726
PRIMER_RIGHT_0_TM=70.180
PRIMER_LEFT_0_GC_PERCENT=75.000
PRIMER_RIGHT_0_GC_PERCENT=54.839
PRIMER_LEFT_0_SELF_ANY_TH=55.63
PRIMER_RIGHT_0_SELF_ANY_TH=14.12
PRIMER_LEFT_0_SELF_END_TH=45.91
PRIMER_RIGHT_0_SELF_END_TH=6.00
PRIMER_LEFT_0_HAIRPIN_TH=74.07
PRIMER_RIGHT_0_HAIRPIN_TH=43.63
PRIMER_LEFT_0_END_STABILITY=5.4000
PRIMER_RIGHT_0_END_STABILITY=4.9600
PRIMER_PAIR_0_COMPL_ANY_TH=34.86
PRIMER_PAIR_0_COMPL_END_TH=44.57
PRIMER_PAIR_0_PRODUCT_SIZE=139
PRIMER_PAIR_0_PRODUCT_TM=85.7
=
PRIMER_LIBERAL_BASE=0
SEQUENCE_ID=Overhang09
SEQUENCE_TEMPLATE=TGCTTTCAATCATGTGGGTGATTTTATAGATTGTCATCACCATAGTAACTGTTTTAACTCACGTCCCCTGTGTGAGAACGCCATGGGCTGTCGGGCTTCGCCAACTTGCTCTCATTTCTA
SEQUENCE_PRIMER=TGCTTTCAATCATGTGGGTGA
SEQUENCE_PRIMER_REVCOMP=ATGAGAGCAAGTTGGCGAAG
SEQUENCE_OVERHANG_LEFT=Z
SEQUENCE_OVERHANG_RIGHT=ATCAG
P3_COMMENT=non-ACGT base in left overhang with PRIMER_LIBERAL_BASE=0
PRIMER_ERROR=Unrecognized base in left overhang
=
SEQUENCE_ID=Overhang10
SEQUENCE_TEMPLATE=TGCTTTCAATCATGTGGGTGATTTTATAGATTGTCATCACCATAGTAACTGTTTTAACTCACGTCCCCTGTGTGAGAACGCCATGGGCTGTCGGGCTTCGCCAACTTGCTCTCATTTCTA
SEQUENCE_PRIMER=TGCTTTCAATCATGTGGGTGA
SEQUENCE_PRIMER_REVCOMP=ATGAGAGCAAGTTGGCGAAG
SEQUENCE_OVERHANG_LEFT=GGCATGCA
SEQUENCE_OVERHANG_RIGHT=Z
P3_COMMENT=non-ACGT base in right overhang with PRIMER_LIBERAL_BASE=0
PRIMER_ERROR=Unrecognized base in right overhang
=
SEQUENCE_ID=Overhang11
SEQUENCE_TEMPLATE=TGCTTTCAATCATGTGGGTGATTTTATAGATTGTCATCACCATAGTAACTGTTTTAACTCACGTCCCCTGTGTGAGAACGCCATGGGCTGTCGGGCTTCGCCAACTTGCTCTCATTTCTA
SEQUENCE_PRIMER=TGCTTTCAATCATGTGGGTGA
SEQUENCE_PRIMER_REVCOMP=ATGAGAGCAAGTTGGCGAAG
SEQUENCE_OVERHANG_RIGHT= C G T C
P3_COMMENT=spaces in right overhang with PRIMER_LIBERAL_BASE=0
PRIMER_ERROR=Unrecognized base in right overhang
=
SEQUENCE_ID=Overhang12
SEQUENCE_TEMPLATE=TGCTTTCAATCATGTGGGTGATTTTATAGATTGTCATCACCATAGTAACTGTTTTAACTCACGTCCCCTGTGTGAGAACGCCATGGGCTGTCGGGCTTCGCCAACTTGCTCTCATTTCTA
SEQUENCE_PRIMER=TGCTTTCAATCATGTGGGTGA
SEQUENCE_PRIMER_REVCOMP=ATGAGAGCAAGTTGGCGAAG
SEQUENCE_OVERHANG_LEFT= A T C G
P3_COMMENT=spaces in left overhang with PRIMER_LIBERAL_BASE=0
PRIMER_ERROR=Unrecognized base in left overhang
=
SEQUENCE_ID=Overhang13
SEQUENCE_TEMPLATE=TGCTTTCAATCATGTGGGTGATTTTATAGATTGTCATCACCATAGTAACTGTTTTAACTCACGTCCCCTGTGTGAGAACGCCATGGGCTGTCGGGCTTCGCCAACTTGCTCTCATTTCTA
SEQUENCE_PRIMER=TGCTTTCAATCATGTGGGTGA
SEQUENCE_PRIMER_REVCOMP=ATGAGAGCAAGTTGGCGAAG
SEQUENCE_OVERHANG_LEFT=
SEQUENCE_OVERHANG_RIGHT=
P3_COMMENT=empty strings for left and right overhangs
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.320921
PRIMER_LEFT_0_PENALTY=2.874014
PRIMER_RIGHT_0_PENALTY=1.446907
PRIMER_LEFT_0_SEQUENCE=TGCTTTCAATCATGTGGGTGA
PRIMER_RIGHT_0_SEQUENCE=ATGAGAGCAAGTTGGCGAAG
PRIMER_LEFT_0=0,21
PRIMER_RIGHT_0=114,20
PRIMER_LEFT_0_TM=58.126
PRIMER_RIGHT_0_TM=58.553
PRIMER_LEFT_0_GC_PERCENT=42.857
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_ANY_TH=4.75
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=39.27
PRIMER_LEFT_0_END_STABILITY=4.0200
PRIMER_RIGHT_0_END_STABILITY=3.7900
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=115
PRIMER_PAIR_0_PRODUCT_TM=82.1
=
SEQUENCE_ID=Overhang14
PRIMER_PICK_LEFT_PRIMER=0
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=1
SEQUENCE_PRIMER_REVCOMP=TAAGGATTTGACTACCAGTCCACTTTAAG
P3_COMMENT=calculate hairpin without using overhangs
PRIMER_LEFT_NUM_RETURNED=0
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_RIGHT_0_PENALTY=9.942466
PRIMER_RIGHT_0_SEQUENCE=TAAGGATTTGACTACCAGTCCACTTTAAG
PRIMER_RIGHT_0=939,29
PRIMER_RIGHT_0_TM=60.942
PRIMER_RIGHT_0_GC_PERCENT=37.931
PRIMER_RIGHT_0_SELF_ANY_TH=3.97
PRIMER_RIGHT_0_SELF_ANY_STUCT=Tm: 4.0&deg;C  dG: -3087 cal/mol  dH: -68200 cal/mol  dS: -210 cal/mol*K\n5' TAAGGATTTGACTACCAGTCCACTTTAAG 3'\n            ||||   ||||\n3' GAATTTCACCTGACCATCAGTTTAGGAAT 5'\n
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=44.44
PRIMER_RIGHT_0_HAIRPIN_STUCT=Tm: 44.4&deg;C  dG: -595 cal/mol  dH: -25400 cal/mol  dS: -80 cal/mol*K\n5' TAAGGATTTGACTAU+2510\n            |||| C\n3' GAATTTCACCTGACU+2518\n
PRIMER_RIGHT_0_END_STABILITY=1.8500
=
SEQUENCE_ID=Overhang15
PRIMER_PICK_LEFT_PRIMER=0
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=1
SEQUENCE_PRIMER_REVCOMP=GACTACCAGTCCACTTTAAG
SEQUENCE_OVERHANG_RIGHT=TAAGGATTT
P3_COMMENT=check hairpin calculation with overhang right - same oligo as Overhang14
PRIMER_LEFT_NUM_RETURNED=0
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_RIGHT_0_PENALTY=7.318716
PRIMER_RIGHT_0_PROBLEMS= Temperature too low;
PRIMER_RIGHT_0_SEQUENCE=TAAGGATTTGACTACCAGTCCACTTTAAG
PRIMER_RIGHT_0=939,20
PRIMER_RIGHT_0_TM=52.681
PRIMER_RIGHT_0_GC_PERCENT=45.000
PRIMER_RIGHT_0_SELF_ANY_TH=3.97
PRIMER_RIGHT_0_SELF_ANY_STUCT=Tm: 4.0&deg;C  dG: -3087 cal/mol  dH: -68200 cal/mol  dS: -210 cal/mol*K\n5' TAAGGATTTGACTACCAGTCCACTTTAAG 3'\n            ||||   ||||\n3' GAATTTCACCTGACCATCAGTTTAGGAAT 5'\n
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=44.44
PRIMER_RIGHT_0_HAIRPIN_STUCT=Tm: 44.4&deg;C  dG: -595 cal/mol  dH: -25400 cal/mol  dS: -80 cal/mol*K\n5' TAAGGATTTGACTAU+2510\n            |||| C\n3' GAATTTCACCTGACU+2518\n
PRIMER_RIGHT_0_END_STABILITY=1.8500
=
SEQUENCE_ID=Overhang16
PRIMER_PICK_LEFT_PRIMER=0
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=0
SEQUENCE_PRIMER_REVCOMP=GACTACCAGTCCACTTTAAG
SEQUENCE_OVERHANG_RIGHT=TAAGGATTT
P3_COMMENT=same as Overhang15 but with PRIMER_SECONDARY_STRUCTURE_ALIGNMENT=0
PRIMER_LEFT_NUM_RETURNED=0
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_RIGHT_0_PENALTY=7.318716
PRIMER_RIGHT_0_PROBLEMS= Temperature too low;
PRIMER_RIGHT_0_SEQUENCE=TAAGGATTTGACTACCAGTCCACTTTAAG
PRIMER_RIGHT_0=939,20
PRIMER_RIGHT_0_TM=52.681
PRIMER_RIGHT_0_GC_PERCENT=45.000
PRIMER_RIGHT_0_SELF_ANY_TH=3.97
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=44.44
PRIMER_RIGHT_0_END_STABILITY=1.8500
=
SEQUENCE_ID=Overhang17
PRIMER_LIBERAL_BASE=1
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
SEQUENCE_TEMPLATE=TGCTTTCAATCATGTGGGTGATTTTATAGATTGTCATCACCATAGTAACTGTTTTAACTCACGTCCCCTGTGTGAGAACGCCATGGGCTGTCGGGCTTCGCCAACTTGCTCTCATTTCTA
SEQUENCE_PRIMER=TGCTTTCAATCATGTGGGTGA
SEQUENCE_PRIMER_REVCOMP=ATGAGAGCAAGTTGGCGAAG
SEQUENCE_OVERHANG_LEFT=ATZZCG
SEQUENCE_OVERHANG_RIGHT=ATZZCC
P3_COMMENT=PRIMER_LIBERAL_BASE=1 with overhangs containing non-ACGT base pairs
PRIMER_ERROR=Unrecognized base in left overhang
=
SEQUENCE_ID=Overhang18
PRIMER_TASK=generic
PRIMER_NUM_RETURN=3
PRIMER_LIBERAL_BASE=0
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=1
SEQUENCE_TEMPLATE=TGCTTTCAATCATGTGGGTGATTTTATAGATTGTCATCACCATAGTAACTGTTTTAACTCACGTCCCCTGTGTGAGAACGCCATGGGCTGTCGGGCTTCGCCAACTTGCTCTCATTTCTA
SEQUENCE_OVERHANG_LEFT=ATG
SEQUENCE_OVERHANG_RIGHT=TAA
P3_COMMENT=PRIMER_TASK=generic with overhangs
PRIMER_LEFT_EXPLAIN=considered 1342, GC content failed 7, low tm 643, high tm 321, high hairpin stability 45, ok 326
PRIMER_RIGHT_EXPLAIN=considered 1342, low tm 285, high tm 799, high hairpin stability 7, ok 251
PRIMER_INTERNAL_EXPLAIN=considered 985, low tm 634, high tm 148, ok 203
PRIMER_PAIR_EXPLAIN=considered 3489, unacceptable product size 2177, no internal oligo 1308, ok 4
PRIMER_LEFT_NUM_RETURNED=3
PRIMER_RIGHT_NUM_RETURNED=3
PRIMER_INTERNAL_NUM_RETURNED=3
PRIMER_PAIR_NUM_RETURNED=3
PRIMER_PAIR_0_PENALTY=2.124196
PRIMER_LEFT_0_PENALTY=0.677288
PRIMER_RIGHT_0_PENALTY=1.446907
PRIMER_INTERNAL_0_PENALTY=0.385767
PRIMER_LEFT_0_SEQUENCE=ATGTAACTCACGTCCCCTGTGTG
PRIMER_RIGHT_0_SEQUENCE=TAAATGAGAGCAAGTTGGCGAAG
PRIMER_INTERNAL_0_SEQUENCE=GAACGCCATGGGCTGTCGGG
PRIMER_LEFT_0=54,20
PRIMER_RIGHT_0=114,20
PRIMER_INTERNAL_0=75,20
PRIMER_LEFT_0_TM=59.323
PRIMER_RIGHT_0_TM=58.553
PRIMER_INTERNAL_0_TM=60.386
PRIMER_LEFT_0_GC_PERCENT=55.000
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_INTERNAL_0_GC_PERCENT=70.000
PRIMER_INTERNAL_0_SELF_ANY_TH=12.04
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_ANY_TH=4.75
PRIMER_INTERNAL_0_SELF_END_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=38.48
PRIMER_RIGHT_0_HAIRPIN_TH=39.27
PRIMER_INTERNAL_0_HAIRPIN_TH=44.17
PRIMER_LEFT_0_END_STABILITY=3.8200
PRIMER_RIGHT_0_END_STABILITY=3.7900
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=67
PRIMER_PAIR_0_PRODUCT_TM=82.1
PRIMER_PAIR_1_PENALTY=2.409199
PRIMER_LEFT_1_PENALTY=0.600255
PRIMER_RIGHT_1_PENALTY=1.808945
PRIMER_INTERNAL_1_PENALTY=1.334653
PRIMER_LEFT_1_SEQUENCE=ATGCACGTCCCCTGTGTGAGAAC
PRIMER_RIGHT_1_SEQUENCE=TAAAGAAATGAGAGCAAGTTGGCG
PRIMER_INTERNAL_1_SEQUENCE=GCCATGGGCTGTCGGGCTT
PRIMER_LEFT_1=59,20
PRIMER_RIGHT_1=118,21
PRIMER_INTERNAL_1=79,19
PRIMER_LEFT_1_TM=60.600
PRIMER_RIGHT_1_TM=59.191
PRIMER_INTERNAL_1_TM=59.665
PRIMER_LEFT_1_GC_PERCENT=60.000
PRIMER_RIGHT_1_GC_PERCENT=47.619
PRIMER_INTERNAL_1_GC_PERCENT=68.421
PRIMER_INTERNAL_1_SELF_ANY_TH=12.04
PRIMER_LEFT_1_SELF_ANY_TH=0.00
PRIMER_RIGHT_1_SELF_ANY_TH=4.75
PRIMER_INTERNAL_1_SELF_END_TH=0.00
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=39.53
PRIMER_RIGHT_1_HAIRPIN_TH=39.27
PRIMER_INTERNAL_1_HAIRPIN_TH=42.85
PRIMER_LEFT_1_END_STABILITY=3.0100
PRIMER_RIGHT_1_END_STABILITY=5.6900
PRIMER_PAIR_1_COMPL_ANY_TH=0.00
PRIMER_PAIR_1_COMPL_END_TH=0.00
PRIMER_PAIR_1_PRODUCT_SIZE=66
PRIMER_PAIR_1_PRODUCT_TM=82.4
PRIMER_PAIR_2_PENALTY=2.483831
PRIMER_LEFT_2_PENALTY=0.677288
PRIMER_RIGHT_2_PENALTY=1.806542
PRIMER_INTERNAL_2_PENALTY=0.385767
PRIMER_LEFT_2_SEQUENCE=ATGTAACTCACGTCCCCTGTGTG
PRIMER_RIGHT_2_SEQUENCE=TAAGAAATGAGAGCAAGTTGGCGA
PRIMER_INTERNAL_2_SEQUENCE=GAACGCCATGGGCTGTCGGG
PRIMER_LEFT_2=54,20
PRIMER_RIGHT_2=117,21
PRIMER_INTERNAL_2=75,20
PRIMER_LEFT_2_TM=59.323
PRIMER_RIGHT_2_TM=59.193
PRIMER_INTERNAL_2_TM=60.386
PRIMER_LEFT_2_GC_PERCENT=55.000
PRIMER_RIGHT_2_GC_PERCENT=47.619
PRIMER_INTERNAL_2_GC_PERCENT=70.000
PRIMER_INTERNAL_2_SELF_ANY_TH=12.04
PRIMER_LEFT_2_SELF_ANY_TH=0.00
PRIMER_RIGHT_2_SELF_ANY_TH=4.75
PRIMER_INTERNAL_2_SELF_END_TH=0.00
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=0.00
PRIMER_LEFT_2_HAIRPIN_TH=38.48
PRIMER_RIGHT_2_HAIRPIN_TH=39.27
PRIMER_INTERNAL_2_HAIRPIN_TH=44.17
PRIMER_LEFT_2_END_STABILITY=3.8200
PRIMER_RIGHT_2_END_STABILITY=5.5400
PRIMER_PAIR_2_COMPL_ANY_TH=0.00
PRIMER_PAIR_2_COMPL_END_TH=0.00
PRIMER_PAIR_2_PRODUCT_SIZE=70
PRIMER_PAIR_2_PRODUCT_TM=82.1
=
SEQUENCE_ID=Overhang19
PRIMER_TASK=pick_primer_list
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
SEQUENCE_TEMPLATE=TGCTTTCAATCATGTGGGTGATTTTATAGATTGTCATCACCATAGTAACTGTTTTAACTCACGTCCCCTGTGTGAGAACGCCATGGGCTGTCGGGCTTCGCCAACTTGCTCTCATTTCTA
SEQUENCE_OVERHANG_LEFT=ATCCC
SEQUENCE_OVERHANG_RIGHT=ATTAGTC
P3_COMMENT=PRIMER_TASK=pick_primer_list with overhangs
PRIMER_LEFT_EXPLAIN=considered 2123, GC content failed 7, low tm 790, high tm 836, high hairpin stability 153, ok 337
PRIMER_RIGHT_EXPLAIN=considered 2123, GC content failed 7, low tm 790, high tm 836, high hairpin stability 207, ok 283
PRIMER_PAIR_EXPLAIN=considered 0, ok 0
PRIMER_LEFT_NUM_RETURNED=3
PRIMER_RIGHT_NUM_RETURNED=3
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_LEFT_0_PENALTY=0.600255
PRIMER_RIGHT_0_PENALTY=1.003355
PRIMER_LEFT_0_SEQUENCE=ATCCCCACGTCCCCTGTGTGAGAAC
PRIMER_RIGHT_0_SEQUENCE=ATTAGTCATGGCGTTCTCACACAGGG
PRIMER_LEFT_0=59,20
PRIMER_RIGHT_0=83,19
PRIMER_LEFT_0_TM=60.600
PRIMER_RIGHT_0_TM=60.003
PRIMER_LEFT_0_GC_PERCENT=60.000
PRIMER_RIGHT_0_GC_PERCENT=57.895
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=35.66
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=3.0100
PRIMER_RIGHT_0_END_STABILITY=4.4500
PRIMER_LEFT_1_PENALTY=0.677288
PRIMER_RIGHT_1_PENALTY=1.300971
PRIMER_LEFT_1_SEQUENCE=ATCCCTAACTCACGTCCCCTGTGTG
PRIMER_RIGHT_1_SEQUENCE=ATTAGTCCCCATGGCGTTCTCACACAG
PRIMER_LEFT_1=54,20
PRIMER_RIGHT_1=86,20
PRIMER_LEFT_1_TM=59.323
PRIMER_RIGHT_1_TM=61.301
PRIMER_LEFT_1_GC_PERCENT=55.000
PRIMER_RIGHT_1_GC_PERCENT=60.000
PRIMER_LEFT_1_SELF_ANY_TH=0.00
PRIMER_RIGHT_1_SELF_ANY_TH=6.09
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=38.48
PRIMER_RIGHT_1_HAIRPIN_TH=0.00
PRIMER_LEFT_1_END_STABILITY=3.8200
PRIMER_RIGHT_1_END_STABILITY=3.6600
PRIMER_LEFT_2_PENALTY=1.003355
PRIMER_RIGHT_2_PENALTY=1.300971
PRIMER_LEFT_2_SEQUENCE=ATCCCCCCTGTGTGAGAACGCCAT
PRIMER_RIGHT_2_SEQUENCE=ATTAGTCCCATGGCGTTCTCACACAGG
PRIMER_LEFT_2=65,19
PRIMER_RIGHT_2=85,20
PRIMER_LEFT_2_TM=60.003
PRIMER_RIGHT_2_TM=61.301
PRIMER_LEFT_2_GC_PERCENT=57.895
PRIMER_RIGHT_2_GC_PERCENT=60.000
PRIMER_LEFT_2_SELF_ANY_TH=0.00
PRIMER_RIGHT_2_SELF_ANY_TH=6.09
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=0.00
PRIMER_LEFT_2_HAIRPIN_TH=0.00
PRIMER_RIGHT_2_HAIRPIN_TH=0.00
PRIMER_LEFT_2_END_STABILITY=4.4000
PRIMER_RIGHT_2_END_STABILITY=4.0000
=
SEQUENCE_ID=Overhang20
PRIMER_TASK=pick_sequencing_primers
PRIMER_EXPLAIN_FLAG=0
SEQUENCE_TEMPLATE=TGCTTTCAATCATGTGGGTGATTTTATAGATTGTCATCACCATAGTAACTGTTTTAACTCACGTCCCCTGTGTGAGAACGCCATGGGCTGTCGGGCTTCGCCAACTTGCTCTCATTTCTA
SEQUENCE_OVERHANG_LEFT=ATCCC
SEQUENCE_OVERHANG_RIGHT=ATTAGTC
P3_COMMENT=PRIMER_TASK=pick_sequencing_primers with overhangs
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_LEFT_0_PENALTY=2.874014
PRIMER_RIGHT_0_PENALTY=1.446907
PRIMER_LEFT_0_SEQUENCE=ATCCCTGCTTTCAATCATGTGGGTGA
PRIMER_RIGHT_0_SEQUENCE=ATTAGTCATGAGAGCAAGTTGGCGAAG
PRIMER_LEFT_0=0,21
PRIMER_RIGHT_0=114,20
PRIMER_LEFT_0_TM=58.126
PRIMER_RIGHT_0_TM=58.553
PRIMER_LEFT_0_GC_PERCENT=42.857
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_ANY_TH=4.75
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=34.14
PRIMER_RIGHT_0_HAIRPIN_TH=39.27
PRIMER_LEFT_0_END_STABILITY=4.0200
PRIMER_RIGHT_0_END_STABILITY=3.7900
=
SEQUENCE_ID=Overhang21
PRIMER_TASK=pick_cloning_primers
PRIMER_EXPLAIN_FLAG=1
SEQUENCE_TEMPLATE=TGCTTTCAATCATGTGGGTGATTTTATAGATTGTCATCACCATAGTAACTGTTTTAACTCACGTCCCCTGTGTGAGAACGCCATGGGCTGTCGGGCTTCGCCAACTTGCTCTCATTTCTA
SEQUENCE_INCLUDED_REGION=25,90
SEQUENCE_OVERHANG_LEFT=ATCCC
SEQUENCE_OVERHANG_RIGHT=ATTAGTC
P3_COMMENT=PRIMER_TASK=pick_cloning_primers with overhangs
PRIMER_LEFT_EXPLAIN=considered 22, low tm 13, ok 22
PRIMER_RIGHT_EXPLAIN=considered 22, low tm 5, high tm 14, high hairpin stability 15, ok 22
PRIMER_PAIR_EXPLAIN=considered 3, ok 3
PRIMER_LEFT_NUM_RETURNED=3
PRIMER_RIGHT_NUM_RETURNED=3
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=3
PRIMER_PAIR_0_PENALTY=10.375356
PRIMER_LEFT_0_PENALTY=8.928448
PRIMER_RIGHT_0_PENALTY=1.446907
PRIMER_LEFT_0_SEQUENCE=ATCCCATAGATTGTCATCACCATAGTAACTGT
PRIMER_RIGHT_0_SEQUENCE=ATTAGTCATGAGAGCAAGTTGGCGAAG
PRIMER_LEFT_0=25,27
PRIMER_RIGHT_0=114,20
PRIMER_LEFT_0_TM=58.072
PRIMER_RIGHT_0_TM=58.553
PRIMER_LEFT_0_GC_PERCENT=33.333
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_ANY_TH=4.75
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=39.27
PRIMER_LEFT_0_END_STABILITY=3.5500
PRIMER_RIGHT_0_END_STABILITY=3.7900
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=102
PRIMER_PAIR_0_PRODUCT_TM=81.8
PRIMER_PAIR_1_PENALTY=10.798901
PRIMER_LEFT_1_PENALTY=9.351993
PRIMER_RIGHT_1_PENALTY=1.446907
PRIMER_LEFT_1_PROBLEMS= Temperature too low;
PRIMER_LEFT_1_SEQUENCE=ATCCCATAGATTGTCATCACCATAGTAACTG
PRIMER_RIGHT_1_SEQUENCE=ATTAGTCATGAGAGCAAGTTGGCGAAG
PRIMER_LEFT_1=25,26
PRIMER_RIGHT_1=114,20
PRIMER_LEFT_1_TM=56.648
PRIMER_RIGHT_1_TM=58.553
PRIMER_LEFT_1_GC_PERCENT=34.615
PRIMER_RIGHT_1_GC_PERCENT=50.000
PRIMER_LEFT_1_SELF_ANY_TH=0.00
PRIMER_RIGHT_1_SELF_ANY_TH=4.75
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=0.00
PRIMER_RIGHT_1_HAIRPIN_TH=39.27
PRIMER_LEFT_1_END_STABILITY=3.1600
PRIMER_RIGHT_1_END_STABILITY=3.7900
PRIMER_PAIR_1_COMPL_ANY_TH=0.00
PRIMER_PAIR_1_COMPL_END_TH=0.00
PRIMER_PAIR_1_PRODUCT_SIZE=102
PRIMER_PAIR_1_PRODUCT_TM=81.8
PRIMER_PAIR_2_PENALTY=10.859739
PRIMER_LEFT_2_PENALTY=9.412832
PRIMER_RIGHT_2_PENALTY=1.446907
PRIMER_LEFT_2_SEQUENCE=ATCCCATAGATTGTCATCACCATAGTAACTGTT
PRIMER_RIGHT_2_SEQUENCE=ATTAGTCATGAGAGCAAGTTGGCGAAG
PRIMER_LEFT_2=25,28
PRIMER_RIGHT_2=114,20
PRIMER_LEFT_2_TM=58.587
PRIMER_RIGHT_2_TM=58.553
PRIMER_LEFT_2_GC_PERCENT=32.143
PRIMER_RIGHT_2_GC_PERCENT=50.000
PRIMER_LEFT_2_SELF_ANY_TH=0.00
PRIMER_RIGHT_2_SELF_ANY_TH=4.75
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=0.00
PRIMER_LEFT_2_HAIRPIN_TH=0.00
PRIMER_RIGHT_2_HAIRPIN_TH=39.27
PRIMER_LEFT_2_END_STABILITY=3.1600
PRIMER_RIGHT_2_END_STABILITY=3.7900
PRIMER_PAIR_2_COMPL_ANY_TH=0.00
PRIMER_PAIR_2_COMPL_END_TH=0.00
PRIMER_PAIR_2_PRODUCT_SIZE=102
PRIMER_PAIR_2_PRODUCT_TM=81.8
=
SEQUENCE_ID=Overhang22
PRIMER_TASK=pick_discriminative_primers
SEQUENCE_TEMPLATE=TGCTTTCAATCATGTGGGTGATTTTATAGATTGTCATCACCATAGTAACTGTTTTAACTCACGTCCCCTGTGTGAGAACGCCATGGGCTGTCGGGCTTCGCCAACTTGCTCTCATTTCTA
SEQUENCE_TARGET=25,60
SEQUENCE_OVERHANG_LEFT=ATCCC
SEQUENCE_OVERHANG_RIGHT=ATTAGTC
P3_COMMENT=PRIMER_TASK=pick_discriminative_primers with overhangs
PRIMER_LEFT_EXPLAIN=considered 12, low tm 10, high hairpin stability 7, ok 12
PRIMER_RIGHT_EXPLAIN=considered 22, low tm 1, high tm 19, high hairpin stability 21, ok 22
PRIMER_PAIR_EXPLAIN=considered 4, ok 4
PRIMER_LEFT_NUM_RETURNED=3
PRIMER_RIGHT_NUM_RETURNED=3
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=3
PRIMER_PAIR_0_PENALTY=10.132060
PRIMER_LEFT_0_PENALTY=5.221682
PRIMER_RIGHT_0_PENALTY=4.910378
PRIMER_RIGHT_0_PROBLEMS= Hairpin stability too high;
PRIMER_LEFT_0_SEQUENCE=ATCCCTGCTTTCAATCATGTGGGTGATTTT
PRIMER_RIGHT_0_SEQUENCE=ATTAGTCGCGAAGCCCGACAGCC
PRIMER_LEFT_0=0,25
PRIMER_RIGHT_0=100,16
PRIMER_LEFT_0_TM=60.222
PRIMER_RIGHT_0_TM=60.910
PRIMER_LEFT_0_GC_PERCENT=36.000
PRIMER_RIGHT_0_GC_PERCENT=75.000
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_ANY_TH=12.06
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=44.23
PRIMER_RIGHT_0_HAIRPIN_TH=62.23
PRIMER_LEFT_0_END_STABILITY=1.8200
PRIMER_RIGHT_0_END_STABILITY=4.8500
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=113
PRIMER_PAIR_0_PRODUCT_TM=81.6
PRIMER_PAIR_1_PENALTY=10.150499
PRIMER_LEFT_1_PENALTY=5.240121
PRIMER_RIGHT_1_PENALTY=4.910378
PRIMER_RIGHT_1_PROBLEMS= Hairpin stability too high;
PRIMER_LEFT_1_SEQUENCE=ATCCCGCTTTCAATCATGTGGGTGATTTT
PRIMER_RIGHT_1_SEQUENCE=ATTAGTCGCGAAGCCCGACAGCC
PRIMER_LEFT_1=1,24
PRIMER_RIGHT_1=100,16
PRIMER_LEFT_1_TM=58.760
PRIMER_RIGHT_1_TM=60.910
PRIMER_LEFT_1_GC_PERCENT=37.500
PRIMER_RIGHT_1_GC_PERCENT=75.000
PRIMER_LEFT_1_SELF_ANY_TH=9.82
PRIMER_RIGHT_1_SELF_ANY_TH=12.06
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=44.23
PRIMER_RIGHT_1_HAIRPIN_TH=62.23
PRIMER_LEFT_1_END_STABILITY=1.8200
PRIMER_RIGHT_1_END_STABILITY=4.8500
PRIMER_PAIR_1_COMPL_ANY_TH=0.00
PRIMER_PAIR_1_COMPL_END_TH=0.00
PRIMER_PAIR_1_PRODUCT_SIZE=112
PRIMER_PAIR_1_PRODUCT_TM=81.7
PRIMER_PAIR_2_PENALTY=11.865677
PRIMER_LEFT_2_PENALTY=5.221682
PRIMER_RIGHT_2_PENALTY=6.643995
PRIMER_RIGHT_2_PROBLEMS= Temperature too high; Hairpin stability too high;
PRIMER_LEFT_2_SEQUENCE=ATCCCTGCTTTCAATCATGTGGGTGATTTT
PRIMER_RIGHT_2_SEQUENCE=ATTAGTCGGCGAAGCCCGACAGCC
PRIMER_LEFT_2=0,25
PRIMER_RIGHT_2=101,17
PRIMER_LEFT_2_TM=60.222
PRIMER_RIGHT_2_TM=63.644
PRIMER_LEFT_2_GC_PERCENT=36.000
PRIMER_RIGHT_2_GC_PERCENT=76.471
PRIMER_LEFT_2_SELF_ANY_TH=0.00
PRIMER_RIGHT_2_SELF_ANY_TH=28.51
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=12.56
PRIMER_LEFT_2_HAIRPIN_TH=44.23
PRIMER_RIGHT_2_HAIRPIN_TH=80.01
PRIMER_LEFT_2_END_STABILITY=1.8200
PRIMER_RIGHT_2_END_STABILITY=4.8500
PRIMER_PAIR_2_COMPL_ANY_TH=0.00
PRIMER_PAIR_2_COMPL_END_TH=0.00
PRIMER_PAIR_2_PRODUCT_SIZE=114
PRIMER_PAIR_2_PRODUCT_TM=81.9
=
SEQUENCE_ID=Overhang23
PRIMER_TASK=generic
PRIMER_NUM_RETURN=3
PRIMER_LIBERAL_BASE=0
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_MAX_SIZE=27
SEQUENCE_TEMPLATE=TGCTTTCAATCATGTGGGTGATTTTATAGATTGTCATCACCATAGTAACTGTTTTAACTCACGTCCCCTGTGTGAGAACGCCATGGGCTGTCGGGCTTCGCCAACTTGCTCTCATTTCTA
SEQUENCE_OVERHANG_LEFT=AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
SEQUENCE_OVERHANG_RIGHT=CCCCCCCCCCCCCCCCCCCCCCCCCCCCCC
P3_COMMENT=PRIMER_TASK=generic with huge overhangs
PRIMER_LEFT_EXPLAIN=considered 854, GC content failed 7, low tm 635, high tm 66, high hairpin stability 1, ok 145
PRIMER_RIGHT_EXPLAIN=considered 854, low tm 285, high tm 353, high hairpin stability 4, ok 212
PRIMER_PAIR_EXPLAIN=considered 418, unacceptable product size 412, ok 6
PRIMER_LEFT_NUM_RETURNED=3
PRIMER_RIGHT_NUM_RETURNED=3
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=3
PRIMER_PAIR_0_PENALTY=2.124196
PRIMER_LEFT_0_PENALTY=0.677288
PRIMER_RIGHT_0_PENALTY=1.446907
PRIMER_LEFT_0_SEQUENCE=AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATAACTCACGTCCCCTGTGTG
PRIMER_RIGHT_0_SEQUENCE=CCCCCCCCCCCCCCCCCCCCCCCCCCCCCCATGAGAGCAAGTTGGCGAAG
PRIMER_LEFT_0=54,20
PRIMER_RIGHT_0=114,20
PRIMER_LEFT_0_TM=59.323
PRIMER_RIGHT_0_TM=58.553
PRIMER_LEFT_0_GC_PERCENT=55.000
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_ANY_TH=4.75
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=38.48
PRIMER_RIGHT_0_HAIRPIN_TH=39.27
PRIMER_LEFT_0_END_STABILITY=3.8200
PRIMER_RIGHT_0_END_STABILITY=3.7900
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=121
PRIMER_PAIR_0_PRODUCT_TM=82.1
PRIMER_PAIR_1_PENALTY=2.483831
PRIMER_LEFT_1_PENALTY=0.677288
PRIMER_RIGHT_1_PENALTY=1.806542
PRIMER_LEFT_1_SEQUENCE=AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATAACTCACGTCCCCTGTGTG
PRIMER_RIGHT_1_SEQUENCE=CCCCCCCCCCCCCCCCCCCCCCCCCCCCCCGAAATGAGAGCAAGTTGGCGA
PRIMER_LEFT_1=54,20
PRIMER_RIGHT_1=117,21
PRIMER_LEFT_1_TM=59.323
PRIMER_RIGHT_1_TM=59.193
PRIMER_LEFT_1_GC_PERCENT=55.000
PRIMER_RIGHT_1_GC_PERCENT=47.619
PRIMER_LEFT_1_SELF_ANY_TH=0.00
PRIMER_RIGHT_1_SELF_ANY_TH=4.75
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=38.48
PRIMER_RIGHT_1_HAIRPIN_TH=39.27
PRIMER_LEFT_1_END_STABILITY=3.8200
PRIMER_RIGHT_1_END_STABILITY=5.5400
PRIMER_PAIR_1_COMPL_ANY_TH=0.00
PRIMER_PAIR_1_COMPL_END_TH=0.00
PRIMER_PAIR_1_PRODUCT_SIZE=124
PRIMER_PAIR_1_PRODUCT_TM=82.1
PRIMER_PAIR_2_PENALTY=2.486233
PRIMER_LEFT_2_PENALTY=0.677288
PRIMER_RIGHT_2_PENALTY=1.808945
PRIMER_LEFT_2_SEQUENCE=AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATAACTCACGTCCCCTGTGTG
PRIMER_RIGHT_2_SEQUENCE=CCCCCCCCCCCCCCCCCCCCCCCCCCCCCCAATGAGAGCAAGTTGGCGAAG
PRIMER_LEFT_2=54,20
PRIMER_RIGHT_2=115,21
PRIMER_LEFT_2_TM=59.323
PRIMER_RIGHT_2_TM=59.191
PRIMER_LEFT_2_GC_PERCENT=55.000
PRIMER_RIGHT_2_GC_PERCENT=47.619
PRIMER_LEFT_2_SELF_ANY_TH=0.00
PRIMER_RIGHT_2_SELF_ANY_TH=4.75
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=0.00
PRIMER_LEFT_2_HAIRPIN_TH=38.48
PRIMER_RIGHT_2_HAIRPIN_TH=39.76
PRIMER_LEFT_2_END_STABILITY=3.8200
PRIMER_RIGHT_2_END_STABILITY=3.7900
PRIMER_PAIR_2_COMPL_ANY_TH=0.00
PRIMER_PAIR_2_COMPL_END_TH=0.00
PRIMER_PAIR_2_PRODUCT_SIZE=122
PRIMER_PAIR_2_PRODUCT_TM=81.9
=
