PRIMER_LOWERCASE_MASKING=0
PRIMER_EXPLAIN_FLAG=1
PRIMER_TASK=generic
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_NUM_RETURN=4
PRIMER_TM_FORMULA=0
PRIMER_SALT_CORRECTIONS=0
SEQUENCE_ID=Pick regular primers Breslauer - Schildkraut
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_LEFT_EXPLAIN=considered 13974, GC content failed 3, low tm 2664, high tm 7479, long poly-x seq 44, ok 3784
PRIMER_RIGHT_EXPLAIN=considered 13974, GC content failed 3, low tm 2880, high tm 7234, long poly-x seq 44, ok 3813
PRIMER_INTERNAL_EXPLAIN=considered 14749, GC content failed 3, low tm 2973, high tm 7694, long poly-x seq 44, ok 4035
PRIMER_PAIR_EXPLAIN=considered 39, unacceptable product size 32, high end compl 2, ok 5
PRIMER_LEFT_NUM_RETURNED=4
PRIMER_RIGHT_NUM_RETURNED=4
PRIMER_INTERNAL_NUM_RETURNED=4
PRIMER_PAIR_NUM_RETURNED=4
PRIMER_PAIR_0_PENALTY=0.013026
PRIMER_LEFT_0_PENALTY=0.006937
PRIMER_RIGHT_0_PENALTY=0.006089
PRIMER_INTERNAL_0_PENALTY=0.187324
PRIMER_LEFT_0_SEQUENCE=gtcctcgaacagctcagtcc
PRIMER_RIGHT_0_SEQUENCE=gcgtttttattgccgttgtt
PRIMER_INTERNAL_0_SEQUENCE=ccacggatgaggaagaagaa
PRIMER_LEFT_0=198,20
PRIMER_RIGHT_0=315,20
PRIMER_INTERNAL_0=250,20
PRIMER_LEFT_0_TM=59.993
PRIMER_RIGHT_0_TM=60.006
PRIMER_INTERNAL_0_TM=60.187
PRIMER_LEFT_0_GC_PERCENT=60.000
PRIMER_RIGHT_0_GC_PERCENT=40.000
PRIMER_INTERNAL_0_GC_PERCENT=50.000
PRIMER_LEFT_0_SELF_ANY=4.00
PRIMER_RIGHT_0_SELF_ANY=2.00
PRIMER_INTERNAL_0_SELF_ANY=2.00
PRIMER_LEFT_0_SELF_END=2.00
PRIMER_RIGHT_0_SELF_END=0.00
PRIMER_INTERNAL_0_SELF_END=0.00
PRIMER_LEFT_0_END_STABILITY=7.6000
PRIMER_RIGHT_0_END_STABILITY=7.0000
PRIMER_PAIR_0_COMPL_ANY=4.00
PRIMER_PAIR_0_COMPL_END=1.00
PRIMER_PAIR_0_PRODUCT_SIZE=118
PRIMER_PAIR_0_PRODUCT_TM=74.3
PRIMER_PAIR_1_PENALTY=0.020508
PRIMER_LEFT_1_PENALTY=0.014419
PRIMER_RIGHT_1_PENALTY=0.006089
PRIMER_INTERNAL_1_PENALTY=0.006937
PRIMER_LEFT_1_SEQUENCE=ttctctggacacagctccct
PRIMER_RIGHT_1_SEQUENCE=gcgtttttattgccgttgtt
PRIMER_INTERNAL_1_SEQUENCE=gtcctcgaacagctcagtcc
PRIMER_LEFT_1=39,20
PRIMER_RIGHT_1=315,20
PRIMER_INTERNAL_1=198,20
PRIMER_LEFT_1_TM=59.986
PRIMER_RIGHT_1_TM=60.006
PRIMER_INTERNAL_1_TM=59.993
PRIMER_LEFT_1_GC_PERCENT=55.000
PRIMER_RIGHT_1_GC_PERCENT=40.000
PRIMER_INTERNAL_1_GC_PERCENT=60.000
PRIMER_LEFT_1_SELF_ANY=4.00
PRIMER_RIGHT_1_SELF_ANY=2.00
PRIMER_INTERNAL_1_SELF_ANY=4.00
PRIMER_LEFT_1_SELF_END=0.00
PRIMER_RIGHT_1_SELF_END=0.00
PRIMER_INTERNAL_1_SELF_END=2.00
PRIMER_LEFT_1_END_STABILITY=9.4000
PRIMER_RIGHT_1_END_STABILITY=7.0000
PRIMER_PAIR_1_COMPL_ANY=3.00
PRIMER_PAIR_1_COMPL_END=0.00
PRIMER_PAIR_1_PRODUCT_SIZE=277
PRIMER_PAIR_1_PRODUCT_TM=78.6
PRIMER_PAIR_2_PENALTY=0.020508
PRIMER_LEFT_2_PENALTY=0.014419
PRIMER_RIGHT_2_PENALTY=0.006089
PRIMER_INTERNAL_2_PENALTY=0.006937
PRIMER_LEFT_2_SEQUENCE=cttctctggacacagctccc
PRIMER_RIGHT_2_SEQUENCE=gcgtttttattgccgttgtt
PRIMER_INTERNAL_2_SEQUENCE=gtcctcgaacagctcagtcc
PRIMER_LEFT_2=38,20
PRIMER_RIGHT_2=315,20
PRIMER_INTERNAL_2=198,20
PRIMER_LEFT_2_TM=59.986
PRIMER_RIGHT_2_TM=60.006
PRIMER_INTERNAL_2_TM=59.993
PRIMER_LEFT_2_GC_PERCENT=60.000
PRIMER_RIGHT_2_GC_PERCENT=40.000
PRIMER_INTERNAL_2_GC_PERCENT=60.000
PRIMER_LEFT_2_SELF_ANY=4.00
PRIMER_RIGHT_2_SELF_ANY=2.00
PRIMER_INTERNAL_2_SELF_ANY=4.00
PRIMER_LEFT_2_SELF_END=0.00
PRIMER_RIGHT_2_SELF_END=0.00
PRIMER_INTERNAL_2_SELF_END=2.00
PRIMER_LEFT_2_END_STABILITY=9.4000
PRIMER_RIGHT_2_END_STABILITY=7.0000
PRIMER_PAIR_2_COMPL_ANY=3.00
PRIMER_PAIR_2_COMPL_END=0.00
PRIMER_PAIR_2_PRODUCT_SIZE=278
PRIMER_PAIR_2_PRODUCT_TM=78.7
PRIMER_PAIR_3_PENALTY=0.032388
PRIMER_LEFT_3_PENALTY=0.032351
PRIMER_RIGHT_3_PENALTY=0.000037
PRIMER_INTERNAL_3_PENALTY=0.077340
PRIMER_LEFT_3_SEQUENCE=gacacacttgctgctttcca
PRIMER_RIGHT_3_SEQUENCE=gtttcttgtacggtggcgat
PRIMER_INTERNAL_3_SEQUENCE=catcttcggataaccgcatt
PRIMER_LEFT_3=609,20
PRIMER_RIGHT_3=868,20
PRIMER_INTERNAL_3=801,20
PRIMER_LEFT_3_TM=60.032
PRIMER_RIGHT_3_TM=60.000
PRIMER_INTERNAL_3_TM=59.923
PRIMER_LEFT_3_GC_PERCENT=50.000
PRIMER_RIGHT_3_GC_PERCENT=50.000
PRIMER_INTERNAL_3_GC_PERCENT=45.000
PRIMER_LEFT_3_SELF_ANY=3.00
PRIMER_RIGHT_3_SELF_ANY=4.00
PRIMER_INTERNAL_3_SELF_ANY=6.00
PRIMER_LEFT_3_SELF_END=0.00
PRIMER_RIGHT_3_SELF_END=2.00
PRIMER_INTERNAL_3_SELF_END=2.00
PRIMER_LEFT_3_END_STABILITY=8.5000
PRIMER_RIGHT_3_END_STABILITY=9.8000
PRIMER_PAIR_3_COMPL_ANY=3.00
PRIMER_PAIR_3_COMPL_END=1.00
PRIMER_PAIR_3_PRODUCT_SIZE=260
PRIMER_PAIR_3_PRODUCT_TM=78.9
=
PRIMER_TM_FORMULA=1
PRIMER_SALT_CORRECTIONS=1
SEQUENCE_ID=Pick regular primers SantaLucia - SantaLucia
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_LEFT_EXPLAIN=considered 13974, GC content failed 3, low tm 9308, high tm 754, high any compl 23, high end compl 41, long poly-x seq 44, ok 3801
PRIMER_RIGHT_EXPLAIN=considered 13974, GC content failed 3, low tm 9554, high tm 738, high any compl 1, high end compl 6, long poly-x seq 44, ok 3628
PRIMER_INTERNAL_EXPLAIN=considered 14749, GC content failed 3, low tm 10018, high tm 754, long poly-x seq 44, ok 3930
PRIMER_PAIR_EXPLAIN=considered 1058, unacceptable product size 1043, high end compl 9, ok 6
PRIMER_LEFT_NUM_RETURNED=4
PRIMER_RIGHT_NUM_RETURNED=4
PRIMER_INTERNAL_NUM_RETURNED=4
PRIMER_PAIR_NUM_RETURNED=4
PRIMER_PAIR_0_PENALTY=0.353282
PRIMER_LEFT_0_PENALTY=0.043363
PRIMER_RIGHT_0_PENALTY=0.309918
PRIMER_INTERNAL_0_PENALTY=1.987429
PRIMER_LEFT_0_SEQUENCE=tggcgcggaagccttaaccg
PRIMER_RIGHT_0_SEQUENCE=ttatgcgcacccccagcacc
PRIMER_INTERNAL_0_SEQUENCE=gccgctcatttcaccgaagcc
PRIMER_LEFT_0=368,20
PRIMER_RIGHT_0=541,20
PRIMER_INTERNAL_0=483,21
PRIMER_LEFT_0_TM=60.043
PRIMER_RIGHT_0_TM=59.690
PRIMER_INTERNAL_0_TM=59.013
PRIMER_LEFT_0_GC_PERCENT=65.000
PRIMER_RIGHT_0_GC_PERCENT=65.000
PRIMER_INTERNAL_0_GC_PERCENT=61.905
PRIMER_LEFT_0_SELF_ANY=5.00
PRIMER_RIGHT_0_SELF_ANY=6.00
PRIMER_INTERNAL_0_SELF_ANY=3.00
PRIMER_LEFT_0_SELF_END=3.00
PRIMER_RIGHT_0_SELF_END=0.00
PRIMER_INTERNAL_0_SELF_END=1.00
PRIMER_LEFT_0_END_STABILITY=4.4400
PRIMER_RIGHT_0_END_STABILITY=5.0100
PRIMER_PAIR_0_COMPL_ANY=4.00
PRIMER_PAIR_0_COMPL_END=1.00
PRIMER_PAIR_0_PRODUCT_SIZE=174
PRIMER_PAIR_0_PRODUCT_TM=77.4
PRIMER_PAIR_1_PENALTY=0.492647
PRIMER_LEFT_1_PENALTY=0.381130
PRIMER_RIGHT_1_PENALTY=0.111517
PRIMER_INTERNAL_1_PENALTY=1.522944
PRIMER_LEFT_1_SEQUENCE=cgaacgccgcgtccatgtca
PRIMER_RIGHT_1_SEQUENCE=ggtggcgattgacctccggc
PRIMER_INTERNAL_1_SEQUENCE=tggaaggggtccaatgggcct
PRIMER_LEFT_1=707,20
PRIMER_RIGHT_1=857,20
PRIMER_INTERNAL_1=742,21
PRIMER_LEFT_1_TM=60.381
PRIMER_RIGHT_1_TM=60.112
PRIMER_INTERNAL_1_TM=59.477
PRIMER_LEFT_1_GC_PERCENT=65.000
PRIMER_RIGHT_1_GC_PERCENT=70.000
PRIMER_INTERNAL_1_GC_PERCENT=61.905
PRIMER_LEFT_1_SELF_ANY=7.00
PRIMER_RIGHT_1_SELF_ANY=4.00
PRIMER_INTERNAL_1_SELF_ANY=7.00
PRIMER_LEFT_1_SELF_END=3.00
PRIMER_RIGHT_1_SELF_END=2.00
PRIMER_INTERNAL_1_SELF_END=5.00
PRIMER_LEFT_1_END_STABILITY=3.5800
PRIMER_RIGHT_1_END_STABILITY=6.1300
PRIMER_PAIR_1_COMPL_ANY=4.00
PRIMER_PAIR_1_COMPL_END=3.00
PRIMER_PAIR_1_PRODUCT_SIZE=151
PRIMER_PAIR_1_PRODUCT_TM=78.2
PRIMER_PAIR_2_PENALTY=0.492766
PRIMER_LEFT_2_PENALTY=0.381250
PRIMER_RIGHT_2_PENALTY=0.111517
PRIMER_INTERNAL_2_PENALTY=1.522944
PRIMER_LEFT_2_SEQUENCE=acgaacgccgcgtccatgtc
PRIMER_RIGHT_2_SEQUENCE=ggtggcgattgacctccggc
PRIMER_INTERNAL_2_SEQUENCE=tggaaggggtccaatgggcct
PRIMER_LEFT_2=706,20
PRIMER_RIGHT_2=857,20
PRIMER_INTERNAL_2=742,21
PRIMER_LEFT_2_TM=60.381
PRIMER_RIGHT_2_TM=60.112
PRIMER_INTERNAL_2_TM=59.477
PRIMER_LEFT_2_GC_PERCENT=65.000
PRIMER_RIGHT_2_GC_PERCENT=70.000
PRIMER_INTERNAL_2_GC_PERCENT=61.905
PRIMER_LEFT_2_SELF_ANY=7.00
PRIMER_RIGHT_2_SELF_ANY=4.00
PRIMER_INTERNAL_2_SELF_ANY=7.00
PRIMER_LEFT_2_SELF_END=1.00
PRIMER_RIGHT_2_SELF_END=2.00
PRIMER_INTERNAL_2_SELF_END=5.00
PRIMER_LEFT_2_END_STABILITY=3.0600
PRIMER_RIGHT_2_END_STABILITY=6.1300
PRIMER_PAIR_2_COMPL_ANY=4.00
PRIMER_PAIR_2_COMPL_END=2.00
PRIMER_PAIR_2_PRODUCT_SIZE=152
PRIMER_PAIR_2_PRODUCT_TM=78.1
PRIMER_PAIR_3_PENALTY=0.557550
PRIMER_LEFT_3_PENALTY=0.381130
PRIMER_RIGHT_3_PENALTY=0.176420
PRIMER_INTERNAL_3_PENALTY=1.522944
PRIMER_LEFT_3_SEQUENCE=cgaacgccgcgtccatgtca
PRIMER_RIGHT_3_SEQUENCE=ttgacctccggccggttcct
PRIMER_INTERNAL_3_SEQUENCE=tggaaggggtccaatgggcct
PRIMER_LEFT_3=707,20
PRIMER_RIGHT_3=849,20
PRIMER_INTERNAL_3=742,21
PRIMER_LEFT_3_TM=60.381
PRIMER_RIGHT_3_TM=59.824
PRIMER_INTERNAL_3_TM=59.477
PRIMER_LEFT_3_GC_PERCENT=65.000
PRIMER_RIGHT_3_GC_PERCENT=65.000
PRIMER_INTERNAL_3_GC_PERCENT=61.905
PRIMER_LEFT_3_SELF_ANY=7.00
PRIMER_RIGHT_3_SELF_ANY=8.00
PRIMER_INTERNAL_3_SELF_ANY=7.00
PRIMER_LEFT_3_SELF_END=3.00
PRIMER_RIGHT_3_SELF_END=0.00
PRIMER_INTERNAL_3_SELF_END=5.00
PRIMER_LEFT_3_END_STABILITY=3.5800
PRIMER_RIGHT_3_END_STABILITY=3.3600
PRIMER_PAIR_3_COMPL_ANY=4.00
PRIMER_PAIR_3_COMPL_END=2.00
PRIMER_PAIR_3_PRODUCT_SIZE=143
PRIMER_PAIR_3_PRODUCT_TM=77.5
=
PRIMER_TM_FORMULA=1
PRIMER_SALT_CORRECTIONS=2
SEQUENCE_ID=Pick regular primers SantaLucia - Owczarzy
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_LEFT_EXPLAIN=considered 13974, GC content failed 3, low tm 9501, high tm 1060, high end compl 3, long poly-x seq 44, ok 3363
PRIMER_RIGHT_EXPLAIN=considered 13974, GC content failed 3, low tm 9794, high tm 1016, long poly-x seq 44, ok 3117
PRIMER_INTERNAL_EXPLAIN=considered 14749, GC content failed 3, low tm 10244, high tm 1060, long poly-x seq 44, ok 3398
PRIMER_PAIR_EXPLAIN=considered 113, unacceptable product size 108, ok 5
PRIMER_LEFT_NUM_RETURNED=4
PRIMER_RIGHT_NUM_RETURNED=4
PRIMER_INTERNAL_NUM_RETURNED=4
PRIMER_PAIR_NUM_RETURNED=4
PRIMER_PAIR_0_PENALTY=0.067561
PRIMER_LEFT_0_PENALTY=0.058197
PRIMER_RIGHT_0_PENALTY=0.009363
PRIMER_INTERNAL_0_PENALTY=1.339571
PRIMER_LEFT_0_SEQUENCE=gctggcgcggaagccttaac
PRIMER_RIGHT_0_SEQUENCE=cgcacccccagcaccttcta
PRIMER_INTERNAL_0_SEQUENCE=gccgctcatttcaccgaagcc
PRIMER_LEFT_0=366,20
PRIMER_RIGHT_0=536,20
PRIMER_INTERNAL_0=483,21
PRIMER_LEFT_0_TM=59.942
PRIMER_RIGHT_0_TM=60.009
PRIMER_INTERNAL_0_TM=59.660
PRIMER_LEFT_0_GC_PERCENT=65.000
PRIMER_RIGHT_0_GC_PERCENT=65.000
PRIMER_INTERNAL_0_GC_PERCENT=61.905
PRIMER_LEFT_0_SELF_ANY=6.00
PRIMER_RIGHT_0_SELF_ANY=2.00
PRIMER_INTERNAL_0_SELF_ANY=3.00
PRIMER_LEFT_0_SELF_END=2.00
PRIMER_RIGHT_0_SELF_END=2.00
PRIMER_INTERNAL_0_SELF_END=1.00
PRIMER_LEFT_0_END_STABILITY=2.0100
PRIMER_RIGHT_0_END_STABILITY=2.1000
PRIMER_PAIR_0_COMPL_ANY=5.00
PRIMER_PAIR_0_COMPL_END=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=171
PRIMER_PAIR_0_PRODUCT_TM=78.0
PRIMER_PAIR_1_PENALTY=0.083042
PRIMER_LEFT_1_PENALTY=0.009363
PRIMER_RIGHT_1_PENALTY=0.073678
PRIMER_INTERNAL_1_PENALTY=0.214552
PRIMER_LEFT_1_SEQUENCE=tagaaggtgctgggggtgcg
PRIMER_RIGHT_1_SEQUENCE=aggcccattggaccccttcc
PRIMER_INTERNAL_1_SEQUENCE=gccatcatcgaagccgtggc
PRIMER_LEFT_1=517,20
PRIMER_RIGHT_1=762,20
PRIMER_INTERNAL_1=684,20
PRIMER_LEFT_1_TM=60.009
PRIMER_RIGHT_1_TM=60.074
PRIMER_INTERNAL_1_TM=59.785
PRIMER_LEFT_1_GC_PERCENT=65.000
PRIMER_RIGHT_1_GC_PERCENT=65.000
PRIMER_INTERNAL_1_GC_PERCENT=65.000
PRIMER_LEFT_1_SELF_ANY=2.00
PRIMER_RIGHT_1_SELF_ANY=7.00
PRIMER_INTERNAL_1_SELF_ANY=4.00
PRIMER_LEFT_1_SELF_END=2.00
PRIMER_RIGHT_1_SELF_END=1.00
PRIMER_INTERNAL_1_SELF_END=4.00
PRIMER_LEFT_1_END_STABILITY=5.3400
PRIMER_RIGHT_1_END_STABILITY=3.4600
PRIMER_PAIR_1_COMPL_ANY=5.00
PRIMER_PAIR_1_COMPL_END=2.00
PRIMER_PAIR_1_PRODUCT_SIZE=246
PRIMER_PAIR_1_PRODUCT_TM=78.3
PRIMER_PAIR_2_PENALTY=0.223915
PRIMER_LEFT_2_PENALTY=0.009363
PRIMER_RIGHT_2_PENALTY=0.214552
PRIMER_INTERNAL_2_PENALTY=2.964100
PRIMER_LEFT_2_SEQUENCE=tagaaggtgctgggggtgcg
PRIMER_RIGHT_2_SEQUENCE=gccacggcttcgatgatggc
PRIMER_INTERNAL_2_SEQUENCE=cggccacttcacagccaatca
PRIMER_LEFT_2=517,20
PRIMER_RIGHT_2=703,20
PRIMER_INTERNAL_2=662,21
PRIMER_LEFT_2_TM=60.009
PRIMER_RIGHT_2_TM=59.785
PRIMER_INTERNAL_2_TM=58.036
PRIMER_LEFT_2_GC_PERCENT=65.000
PRIMER_RIGHT_2_GC_PERCENT=65.000
PRIMER_INTERNAL_2_GC_PERCENT=57.143
PRIMER_LEFT_2_SELF_ANY=2.00
PRIMER_RIGHT_2_SELF_ANY=4.00
PRIMER_INTERNAL_2_SELF_ANY=4.00
PRIMER_LEFT_2_SELF_END=2.00
PRIMER_RIGHT_2_SELF_END=2.00
PRIMER_INTERNAL_2_SELF_END=1.00
PRIMER_LEFT_2_END_STABILITY=5.3400
PRIMER_RIGHT_2_END_STABILITY=4.4000
PRIMER_PAIR_2_COMPL_ANY=4.00
PRIMER_PAIR_2_COMPL_END=1.00
PRIMER_PAIR_2_PRODUCT_SIZE=187
PRIMER_PAIR_2_PRODUCT_TM=76.6
PRIMER_PAIR_3_PENALTY=0.314221
PRIMER_LEFT_3_PENALTY=0.009363
PRIMER_RIGHT_3_PENALTY=0.304858
PRIMER_INTERNAL_3_PENALTY=0.214552
PRIMER_LEFT_3_SEQUENCE=tagaaggtgctgggggtgcg
PRIMER_RIGHT_3_SEQUENCE=ggcccattggaccccttcca
PRIMER_INTERNAL_3_SEQUENCE=gccatcatcgaagccgtggc
PRIMER_LEFT_3=517,20
PRIMER_RIGHT_3=761,20
PRIMER_INTERNAL_3=684,20
PRIMER_LEFT_3_TM=60.009
PRIMER_RIGHT_3_TM=60.305
PRIMER_INTERNAL_3_TM=59.785
PRIMER_LEFT_3_GC_PERCENT=65.000
PRIMER_RIGHT_3_GC_PERCENT=65.000
PRIMER_INTERNAL_3_GC_PERCENT=65.000
PRIMER_LEFT_3_SELF_ANY=2.00
PRIMER_RIGHT_3_SELF_ANY=7.00
PRIMER_INTERNAL_3_SELF_ANY=4.00
PRIMER_LEFT_3_SELF_END=2.00
PRIMER_RIGHT_3_SELF_END=3.00
PRIMER_INTERNAL_3_SELF_END=4.00
PRIMER_LEFT_3_END_STABILITY=5.3400
PRIMER_RIGHT_3_END_STABILITY=3.5300
PRIMER_PAIR_3_COMPL_ANY=5.00
PRIMER_PAIR_3_COMPL_END=3.00
PRIMER_PAIR_3_PRODUCT_SIZE=245
PRIMER_PAIR_3_PRODUCT_TM=78.4
=
PRIMER_TM_FORMULA=0
PRIMER_SALT_CORRECTIONS=0
PRIMER_ANNEALING_TEMP=50.0
SEQUENCE_ID=Pick regular primers with bound fraction Breslauer - Schildkraut
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_LEFT_EXPLAIN=considered 13974, GC content failed 3, low tm 2664, high tm 7479, long poly-x seq 44, ok 3784
PRIMER_RIGHT_EXPLAIN=considered 13974, GC content failed 3, low tm 2880, high tm 7234, long poly-x seq 44, ok 3813
PRIMER_INTERNAL_EXPLAIN=considered 14749, GC content failed 3, low tm 2973, high tm 7694, long poly-x seq 44, ok 4035
PRIMER_PAIR_EXPLAIN=considered 39, unacceptable product size 32, high end compl 2, ok 5
PRIMER_LEFT_NUM_RETURNED=4
PRIMER_RIGHT_NUM_RETURNED=4
PRIMER_INTERNAL_NUM_RETURNED=4
PRIMER_PAIR_NUM_RETURNED=4
PRIMER_PAIR_0_PENALTY=0.013026
PRIMER_LEFT_0_PENALTY=0.006937
PRIMER_RIGHT_0_PENALTY=0.006089
PRIMER_INTERNAL_0_PENALTY=0.187324
PRIMER_LEFT_0_SEQUENCE=gtcctcgaacagctcagtcc
PRIMER_RIGHT_0_SEQUENCE=gcgtttttattgccgttgtt
PRIMER_INTERNAL_0_SEQUENCE=ccacggatgaggaagaagaa
PRIMER_LEFT_0=198,20
PRIMER_RIGHT_0=315,20
PRIMER_INTERNAL_0=250,20
PRIMER_LEFT_0_TM=59.993
PRIMER_RIGHT_0_TM=60.006
PRIMER_INTERNAL_0_TM=60.187
PRIMER_LEFT_0_BOUND=95.116
PRIMER_RIGHT_0_BOUND=96.850
PRIMER_INTERNAL_0_BOUND=95.886
PRIMER_LEFT_0_GC_PERCENT=60.000
PRIMER_RIGHT_0_GC_PERCENT=40.000
PRIMER_INTERNAL_0_GC_PERCENT=50.000
PRIMER_LEFT_0_SELF_ANY=4.00
PRIMER_RIGHT_0_SELF_ANY=2.00
PRIMER_INTERNAL_0_SELF_ANY=2.00
PRIMER_LEFT_0_SELF_END=2.00
PRIMER_RIGHT_0_SELF_END=0.00
PRIMER_INTERNAL_0_SELF_END=0.00
PRIMER_LEFT_0_END_STABILITY=7.6000
PRIMER_RIGHT_0_END_STABILITY=7.0000
PRIMER_PAIR_0_COMPL_ANY=4.00
PRIMER_PAIR_0_COMPL_END=1.00
PRIMER_PAIR_0_PRODUCT_SIZE=118
PRIMER_PAIR_0_PRODUCT_TM=74.3
PRIMER_PAIR_1_PENALTY=0.020508
PRIMER_LEFT_1_PENALTY=0.014419
PRIMER_RIGHT_1_PENALTY=0.006089
PRIMER_INTERNAL_1_PENALTY=0.006937
PRIMER_LEFT_1_SEQUENCE=ttctctggacacagctccct
PRIMER_RIGHT_1_SEQUENCE=gcgtttttattgccgttgtt
PRIMER_INTERNAL_1_SEQUENCE=gtcctcgaacagctcagtcc
PRIMER_LEFT_1=39,20
PRIMER_RIGHT_1=315,20
PRIMER_INTERNAL_1=198,20
PRIMER_LEFT_1_TM=59.986
PRIMER_RIGHT_1_TM=60.006
PRIMER_INTERNAL_1_TM=59.993
PRIMER_LEFT_1_BOUND=95.163
PRIMER_RIGHT_1_BOUND=96.850
PRIMER_INTERNAL_1_BOUND=95.116
PRIMER_LEFT_1_GC_PERCENT=55.000
PRIMER_RIGHT_1_GC_PERCENT=40.000
PRIMER_INTERNAL_1_GC_PERCENT=60.000
PRIMER_LEFT_1_SELF_ANY=4.00
PRIMER_RIGHT_1_SELF_ANY=2.00
PRIMER_INTERNAL_1_SELF_ANY=4.00
PRIMER_LEFT_1_SELF_END=0.00
PRIMER_RIGHT_1_SELF_END=0.00
PRIMER_INTERNAL_1_SELF_END=2.00
PRIMER_LEFT_1_END_STABILITY=9.4000
PRIMER_RIGHT_1_END_STABILITY=7.0000
PRIMER_PAIR_1_COMPL_ANY=3.00
PRIMER_PAIR_1_COMPL_END=0.00
PRIMER_PAIR_1_PRODUCT_SIZE=277
PRIMER_PAIR_1_PRODUCT_TM=78.6
PRIMER_PAIR_2_PENALTY=0.020508
PRIMER_LEFT_2_PENALTY=0.014419
PRIMER_RIGHT_2_PENALTY=0.006089
PRIMER_INTERNAL_2_PENALTY=0.006937
PRIMER_LEFT_2_SEQUENCE=cttctctggacacagctccc
PRIMER_RIGHT_2_SEQUENCE=gcgtttttattgccgttgtt
PRIMER_INTERNAL_2_SEQUENCE=gtcctcgaacagctcagtcc
PRIMER_LEFT_2=38,20
PRIMER_RIGHT_2=315,20
PRIMER_INTERNAL_2=198,20
PRIMER_LEFT_2_TM=59.986
PRIMER_RIGHT_2_TM=60.006
PRIMER_INTERNAL_2_TM=59.993
PRIMER_LEFT_2_BOUND=95.163
PRIMER_RIGHT_2_BOUND=96.850
PRIMER_INTERNAL_2_BOUND=95.116
PRIMER_LEFT_2_GC_PERCENT=60.000
PRIMER_RIGHT_2_GC_PERCENT=40.000
PRIMER_INTERNAL_2_GC_PERCENT=60.000
PRIMER_LEFT_2_SELF_ANY=4.00
PRIMER_RIGHT_2_SELF_ANY=2.00
PRIMER_INTERNAL_2_SELF_ANY=4.00
PRIMER_LEFT_2_SELF_END=0.00
PRIMER_RIGHT_2_SELF_END=0.00
PRIMER_INTERNAL_2_SELF_END=2.00
PRIMER_LEFT_2_END_STABILITY=9.4000
PRIMER_RIGHT_2_END_STABILITY=7.0000
PRIMER_PAIR_2_COMPL_ANY=3.00
PRIMER_PAIR_2_COMPL_END=0.00
PRIMER_PAIR_2_PRODUCT_SIZE=278
PRIMER_PAIR_2_PRODUCT_TM=78.7
PRIMER_PAIR_3_PENALTY=0.032388
PRIMER_LEFT_3_PENALTY=0.032351
PRIMER_RIGHT_3_PENALTY=0.000037
PRIMER_INTERNAL_3_PENALTY=0.077340
PRIMER_LEFT_3_SEQUENCE=gacacacttgctgctttcca
PRIMER_RIGHT_3_SEQUENCE=gtttcttgtacggtggcgat
PRIMER_INTERNAL_3_SEQUENCE=catcttcggataaccgcatt
PRIMER_LEFT_3=609,20
PRIMER_RIGHT_3=868,20
PRIMER_INTERNAL_3=801,20
PRIMER_LEFT_3_TM=60.032
PRIMER_RIGHT_3_TM=60.000
PRIMER_INTERNAL_3_TM=59.923
PRIMER_LEFT_3_BOUND=95.092
PRIMER_RIGHT_3_BOUND=96.074
PRIMER_INTERNAL_3_BOUND=96.234
PRIMER_LEFT_3_GC_PERCENT=50.000
PRIMER_RIGHT_3_GC_PERCENT=50.000
PRIMER_INTERNAL_3_GC_PERCENT=45.000
PRIMER_LEFT_3_SELF_ANY=3.00
PRIMER_RIGHT_3_SELF_ANY=4.00
PRIMER_INTERNAL_3_SELF_ANY=6.00
PRIMER_LEFT_3_SELF_END=0.00
PRIMER_RIGHT_3_SELF_END=2.00
PRIMER_INTERNAL_3_SELF_END=2.00
PRIMER_LEFT_3_END_STABILITY=8.5000
PRIMER_RIGHT_3_END_STABILITY=9.8000
PRIMER_PAIR_3_COMPL_ANY=3.00
PRIMER_PAIR_3_COMPL_END=1.00
PRIMER_PAIR_3_PRODUCT_SIZE=260
PRIMER_PAIR_3_PRODUCT_TM=78.9
=
PRIMER_TM_FORMULA=1
PRIMER_SALT_CORRECTIONS=1
PRIMER_ANNEALING_TEMP=50.0
SEQUENCE_ID=Pick regular primers with bound fraction SantaLucia - SantaLucia
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_LEFT_EXPLAIN=considered 13974, GC content failed 3, low tm 9308, high tm 754, high any compl 23, high end compl 41, long poly-x seq 44, ok 3801
PRIMER_RIGHT_EXPLAIN=considered 13974, GC content failed 3, low tm 9554, high tm 738, high any compl 1, high end compl 6, long poly-x seq 44, ok 3628
PRIMER_INTERNAL_EXPLAIN=considered 14749, GC content failed 3, low tm 10018, high tm 754, long poly-x seq 44, ok 3930
PRIMER_PAIR_EXPLAIN=considered 1058, unacceptable product size 1043, high end compl 9, ok 6
PRIMER_LEFT_NUM_RETURNED=4
PRIMER_RIGHT_NUM_RETURNED=4
PRIMER_INTERNAL_NUM_RETURNED=4
PRIMER_PAIR_NUM_RETURNED=4
PRIMER_PAIR_0_PENALTY=0.353282
PRIMER_LEFT_0_PENALTY=0.043363
PRIMER_RIGHT_0_PENALTY=0.309918
PRIMER_INTERNAL_0_PENALTY=1.987429
PRIMER_LEFT_0_SEQUENCE=tggcgcggaagccttaaccg
PRIMER_RIGHT_0_SEQUENCE=ttatgcgcacccccagcacc
PRIMER_INTERNAL_0_SEQUENCE=gccgctcatttcaccgaagcc
PRIMER_LEFT_0=368,20
PRIMER_RIGHT_0=541,20
PRIMER_INTERNAL_0=483,21
PRIMER_LEFT_0_TM=60.043
PRIMER_RIGHT_0_TM=59.690
PRIMER_INTERNAL_0_TM=59.013
PRIMER_LEFT_0_BOUND=97.837
PRIMER_RIGHT_0_BOUND=97.323
PRIMER_INTERNAL_0_BOUND=97.368
PRIMER_LEFT_0_GC_PERCENT=65.000
PRIMER_RIGHT_0_GC_PERCENT=65.000
PRIMER_INTERNAL_0_GC_PERCENT=61.905
PRIMER_LEFT_0_SELF_ANY=5.00
PRIMER_RIGHT_0_SELF_ANY=6.00
PRIMER_INTERNAL_0_SELF_ANY=3.00
PRIMER_LEFT_0_SELF_END=3.00
PRIMER_RIGHT_0_SELF_END=0.00
PRIMER_INTERNAL_0_SELF_END=1.00
PRIMER_LEFT_0_END_STABILITY=4.4400
PRIMER_RIGHT_0_END_STABILITY=5.0100
PRIMER_PAIR_0_COMPL_ANY=4.00
PRIMER_PAIR_0_COMPL_END=1.00
PRIMER_PAIR_0_PRODUCT_SIZE=174
PRIMER_PAIR_0_PRODUCT_TM=77.4
PRIMER_PAIR_1_PENALTY=0.492647
PRIMER_LEFT_1_PENALTY=0.381130
PRIMER_RIGHT_1_PENALTY=0.111517
PRIMER_INTERNAL_1_PENALTY=1.522944
PRIMER_LEFT_1_SEQUENCE=cgaacgccgcgtccatgtca
PRIMER_RIGHT_1_SEQUENCE=ggtggcgattgacctccggc
PRIMER_INTERNAL_1_SEQUENCE=tggaaggggtccaatgggcct
PRIMER_LEFT_1=707,20
PRIMER_RIGHT_1=857,20
PRIMER_INTERNAL_1=742,21
PRIMER_LEFT_1_TM=60.381
PRIMER_RIGHT_1_TM=60.112
PRIMER_INTERNAL_1_TM=59.477
PRIMER_LEFT_1_BOUND=98.242
PRIMER_RIGHT_1_BOUND=97.865
PRIMER_INTERNAL_1_BOUND=97.090
PRIMER_LEFT_1_GC_PERCENT=65.000
PRIMER_RIGHT_1_GC_PERCENT=70.000
PRIMER_INTERNAL_1_GC_PERCENT=61.905
PRIMER_LEFT_1_SELF_ANY=7.00
PRIMER_RIGHT_1_SELF_ANY=4.00
PRIMER_INTERNAL_1_SELF_ANY=7.00
PRIMER_LEFT_1_SELF_END=3.00
PRIMER_RIGHT_1_SELF_END=2.00
PRIMER_INTERNAL_1_SELF_END=5.00
PRIMER_LEFT_1_END_STABILITY=3.5800
PRIMER_RIGHT_1_END_STABILITY=6.1300
PRIMER_PAIR_1_COMPL_ANY=4.00
PRIMER_PAIR_1_COMPL_END=3.00
PRIMER_PAIR_1_PRODUCT_SIZE=151
PRIMER_PAIR_1_PRODUCT_TM=78.2
PRIMER_PAIR_2_PENALTY=0.492766
PRIMER_LEFT_2_PENALTY=0.381250
PRIMER_RIGHT_2_PENALTY=0.111517
PRIMER_INTERNAL_2_PENALTY=1.522944
PRIMER_LEFT_2_SEQUENCE=acgaacgccgcgtccatgtc
PRIMER_RIGHT_2_SEQUENCE=ggtggcgattgacctccggc
PRIMER_INTERNAL_2_SEQUENCE=tggaaggggtccaatgggcct
PRIMER_LEFT_2=706,20
PRIMER_RIGHT_2=857,20
PRIMER_INTERNAL_2=742,21
PRIMER_LEFT_2_TM=60.381
PRIMER_RIGHT_2_TM=60.112
PRIMER_INTERNAL_2_TM=59.477
PRIMER_LEFT_2_BOUND=98.238
PRIMER_RIGHT_2_BOUND=97.865
PRIMER_INTERNAL_2_BOUND=97.090
PRIMER_LEFT_2_GC_PERCENT=65.000
PRIMER_RIGHT_2_GC_PERCENT=70.000
PRIMER_INTERNAL_2_GC_PERCENT=61.905
PRIMER_LEFT_2_SELF_ANY=7.00
PRIMER_RIGHT_2_SELF_ANY=4.00
PRIMER_INTERNAL_2_SELF_ANY=7.00
PRIMER_LEFT_2_SELF_END=1.00
PRIMER_RIGHT_2_SELF_END=2.00
PRIMER_INTERNAL_2_SELF_END=5.00
PRIMER_LEFT_2_END_STABILITY=3.0600
PRIMER_RIGHT_2_END_STABILITY=6.1300
PRIMER_PAIR_2_COMPL_ANY=4.00
PRIMER_PAIR_2_COMPL_END=2.00
PRIMER_PAIR_2_PRODUCT_SIZE=152
PRIMER_PAIR_2_PRODUCT_TM=78.1
PRIMER_PAIR_3_PENALTY=0.557550
PRIMER_LEFT_3_PENALTY=0.381130
PRIMER_RIGHT_3_PENALTY=0.176420
PRIMER_INTERNAL_3_PENALTY=1.522944
PRIMER_LEFT_3_SEQUENCE=cgaacgccgcgtccatgtca
PRIMER_RIGHT_3_SEQUENCE=ttgacctccggccggttcct
PRIMER_INTERNAL_3_SEQUENCE=tggaaggggtccaatgggcct
PRIMER_LEFT_3=707,20
PRIMER_RIGHT_3=849,20
PRIMER_INTERNAL_3=742,21
PRIMER_LEFT_3_TM=60.381
PRIMER_RIGHT_3_TM=59.824
PRIMER_INTERNAL_3_TM=59.477
PRIMER_LEFT_3_BOUND=98.242
PRIMER_RIGHT_3_BOUND=97.280
PRIMER_INTERNAL_3_BOUND=97.090
PRIMER_LEFT_3_GC_PERCENT=65.000
PRIMER_RIGHT_3_GC_PERCENT=65.000
PRIMER_INTERNAL_3_GC_PERCENT=61.905
PRIMER_LEFT_3_SELF_ANY=7.00
PRIMER_RIGHT_3_SELF_ANY=8.00
PRIMER_INTERNAL_3_SELF_ANY=7.00
PRIMER_LEFT_3_SELF_END=3.00
PRIMER_RIGHT_3_SELF_END=0.00
PRIMER_INTERNAL_3_SELF_END=5.00
PRIMER_LEFT_3_END_STABILITY=3.5800
PRIMER_RIGHT_3_END_STABILITY=3.3600
PRIMER_PAIR_3_COMPL_ANY=4.00
PRIMER_PAIR_3_COMPL_END=2.00
PRIMER_PAIR_3_PRODUCT_SIZE=143
PRIMER_PAIR_3_PRODUCT_TM=77.5
=
PRIMER_TM_FORMULA=0
PRIMER_SALT_CORRECTIONS=0
PRIMER_ANNEALING_TEMP=50.0
PRIMER_MAX_TM=80.0
PRIMER_INTERNAL_MAX_TM=80.0
PRIMER_MIN_TM=50.0
PRIMER_INTERNAL_MIN_TM=50.0
PRIMER_WT_TM_LT=0.0
PRIMER_INTERNAL_WT_TM_LT=0.0
PRIMER_WT_TM_GT=0.0
PRIMER_INTERNAL_WT_TM_GT=0.0
PRIMER_MIN_BOUND=94.0
PRIMER_OPT_BOUND=95.0
PRIMER_MAX_BOUND=96.0
PRIMER_INTERNAL_MIN_BOUND=94.0
PRIMER_INTERNAL_OPT_BOUND=95.0
PRIMER_INTERNAL_MAX_BOUND=96.0
PRIMER_WT_BOUND_LT=1.0
PRIMER_WT_BOUND_GT=1.0
PRIMER_INTERNAL_WT_BOUND_LT=1.0
PRIMER_INTERNAL_WT_BOUND_GT=1.0
SEQUENCE_ID=Pick primers by bound fraction Breslauer - Schildkraut
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_LEFT_EXPLAIN=considered 13974, GC content failed 3, low tm 485, high tm 159, low faction bound 3097, high fraction bound 9525, high any compl 1, high end compl 1, long poly-x seq 44, ok 659
PRIMER_RIGHT_EXPLAIN=considered 13974, GC content failed 3, low tm 579, high tm 159, low faction bound 3191, high fraction bound 9329, high end compl 1, long poly-x seq 44, ok 668
PRIMER_INTERNAL_EXPLAIN=considered 14749, GC content failed 3, low tm 580, high tm 159, low faction bound 3357, high fraction bound 9899, long poly-x seq 44, ok 707
PRIMER_PAIR_EXPLAIN=considered 152, unacceptable product size 142, high end compl 1, ok 9
PRIMER_LEFT_NUM_RETURNED=4
PRIMER_RIGHT_NUM_RETURNED=4
PRIMER_INTERNAL_NUM_RETURNED=4
PRIMER_PAIR_NUM_RETURNED=4
PRIMER_PAIR_0_PENALTY=0.080601
PRIMER_LEFT_0_PENALTY=0.044977
PRIMER_RIGHT_0_PENALTY=0.035625
PRIMER_INTERNAL_0_PENALTY=0.098737
PRIMER_LEFT_0_SEQUENCE=actggtctcccatcgtcaac
PRIMER_RIGHT_0_SEQUENCE=tcttcttcctcatccgtggt
PRIMER_INTERNAL_0_SEQUENCE=tcatggacaccatcatcgag
PRIMER_LEFT_0=106,20
PRIMER_RIGHT_0=268,20
PRIMER_INTERNAL_0=166,20
PRIMER_LEFT_0_TM=59.969
PRIMER_RIGHT_0_TM=59.655
PRIMER_INTERNAL_0_TM=60.496
PRIMER_LEFT_0_BOUND=95.045
PRIMER_RIGHT_0_BOUND=94.964
PRIMER_INTERNAL_0_BOUND=95.099
PRIMER_LEFT_0_GC_PERCENT=55.000
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_INTERNAL_0_GC_PERCENT=50.000
PRIMER_LEFT_0_SELF_ANY=3.00
PRIMER_RIGHT_0_SELF_ANY=2.00
PRIMER_INTERNAL_0_SELF_ANY=5.00
PRIMER_LEFT_0_SELF_END=2.00
PRIMER_RIGHT_0_SELF_END=0.00
PRIMER_INTERNAL_0_SELF_END=3.00
PRIMER_LEFT_0_END_STABILITY=6.7000
PRIMER_RIGHT_0_END_STABILITY=7.6000
PRIMER_PAIR_0_COMPL_ANY=3.00
PRIMER_PAIR_0_COMPL_END=2.00
PRIMER_PAIR_0_PRODUCT_SIZE=163
PRIMER_PAIR_0_PRODUCT_TM=77.9
PRIMER_PAIR_1_PENALTY=0.134361
PRIMER_LEFT_1_PENALTY=0.098737
PRIMER_RIGHT_1_PENALTY=0.035625
PRIMER_INTERNAL_1_PENALTY=0.115917
PRIMER_LEFT_1_SEQUENCE=tcatggacaccatcatcgag
PRIMER_RIGHT_1_SEQUENCE=tcttcttcctcatccgtggt
PRIMER_INTERNAL_1_SEQUENCE=gtcctcgaacagctcagtcc
PRIMER_LEFT_1=166,20
PRIMER_RIGHT_1=268,20
PRIMER_INTERNAL_1=198,20
PRIMER_LEFT_1_TM=60.496
PRIMER_RIGHT_1_TM=59.655
PRIMER_INTERNAL_1_TM=59.993
PRIMER_LEFT_1_BOUND=95.099
PRIMER_RIGHT_1_BOUND=94.964
PRIMER_INTERNAL_1_BOUND=95.116
PRIMER_LEFT_1_GC_PERCENT=50.000
PRIMER_RIGHT_1_GC_PERCENT=50.000
PRIMER_INTERNAL_1_GC_PERCENT=60.000
PRIMER_LEFT_1_SELF_ANY=5.00
PRIMER_RIGHT_1_SELF_ANY=2.00
PRIMER_INTERNAL_1_SELF_ANY=4.00
PRIMER_LEFT_1_SELF_END=3.00
PRIMER_RIGHT_1_SELF_END=0.00
PRIMER_INTERNAL_1_SELF_END=2.00
PRIMER_LEFT_1_END_STABILITY=8.4000
PRIMER_RIGHT_1_END_STABILITY=7.6000
PRIMER_PAIR_1_COMPL_ANY=4.00
PRIMER_PAIR_1_COMPL_END=3.00
PRIMER_PAIR_1_PRODUCT_SIZE=103
PRIMER_PAIR_1_PRODUCT_TM=75.2
PRIMER_PAIR_2_PENALTY=0.140309
PRIMER_LEFT_2_PENALTY=0.049789
PRIMER_RIGHT_2_PENALTY=0.090520
PRIMER_INTERNAL_2_PENALTY=0.060401
PRIMER_LEFT_2_SEQUENCE=gatatcatggaaggggtcca
PRIMER_RIGHT_2_SEQUENCE=aaggacggaaagtttcatcg
PRIMER_INTERNAL_2_SEQUENCE=ccaatcacttgcttccaaca
PRIMER_LEFT_2=735,20
PRIMER_RIGHT_2=960,20
PRIMER_INTERNAL_2=770,20
PRIMER_LEFT_2_TM=59.563
PRIMER_RIGHT_2_TM=59.170
PRIMER_INTERNAL_2_TM=59.691
PRIMER_LEFT_2_BOUND=95.050
PRIMER_RIGHT_2_BOUND=95.091
PRIMER_INTERNAL_2_BOUND=94.940
PRIMER_LEFT_2_GC_PERCENT=50.000
PRIMER_RIGHT_2_GC_PERCENT=45.000
PRIMER_INTERNAL_2_GC_PERCENT=45.000
PRIMER_LEFT_2_SELF_ANY=6.00
PRIMER_RIGHT_2_SELF_ANY=7.00
PRIMER_INTERNAL_2_SELF_ANY=4.00
PRIMER_LEFT_2_SELF_END=3.00
PRIMER_RIGHT_2_SELF_END=3.00
PRIMER_INTERNAL_2_SELF_END=2.00
PRIMER_LEFT_2_END_STABILITY=7.9000
PRIMER_RIGHT_2_END_STABILITY=8.6000
PRIMER_PAIR_2_COMPL_ANY=4.00
PRIMER_PAIR_2_COMPL_END=1.00
PRIMER_PAIR_2_PRODUCT_SIZE=226
PRIMER_PAIR_2_PRODUCT_TM=77.9
PRIMER_PAIR_3_PENALTY=0.147683
PRIMER_LEFT_3_PENALTY=0.044977
PRIMER_RIGHT_3_PENALTY=0.102706
PRIMER_INTERNAL_3_PENALTY=0.035625
PRIMER_LEFT_3_SEQUENCE=actggtctcccatcgtcaac
PRIMER_RIGHT_3_SEQUENCE=tcatcaccaacttcatgcgt
PRIMER_INTERNAL_3_SEQUENCE=accacggatgaggaagaaga
PRIMER_LEFT_3=106,20
PRIMER_RIGHT_3=331,20
PRIMER_INTERNAL_3=249,20
PRIMER_LEFT_3_TM=59.969
PRIMER_RIGHT_3_TM=60.120
PRIMER_INTERNAL_3_TM=59.655
PRIMER_LEFT_3_BOUND=95.045
PRIMER_RIGHT_3_BOUND=95.103
PRIMER_INTERNAL_3_BOUND=94.964
PRIMER_LEFT_3_GC_PERCENT=55.000
PRIMER_RIGHT_3_GC_PERCENT=45.000
PRIMER_INTERNAL_3_GC_PERCENT=50.000
PRIMER_LEFT_3_SELF_ANY=3.00
PRIMER_RIGHT_3_SELF_ANY=4.00
PRIMER_INTERNAL_3_SELF_ANY=2.00
PRIMER_LEFT_3_SELF_END=2.00
PRIMER_RIGHT_3_SELF_END=2.00
PRIMER_INTERNAL_3_SELF_END=0.00
PRIMER_LEFT_3_END_STABILITY=6.7000
PRIMER_RIGHT_3_END_STABILITY=9.9000
PRIMER_PAIR_3_COMPL_ANY=4.00
PRIMER_PAIR_3_COMPL_END=2.00
PRIMER_PAIR_3_PRODUCT_SIZE=226
PRIMER_PAIR_3_PRODUCT_TM=77.2
=
PRIMER_TM_FORMULA=1
PRIMER_SALT_CORRECTIONS=1
PRIMER_ANNEALING_TEMP=50.0
PRIMER_MAX_TM=80.0
PRIMER_INTERNAL_MAX_TM=80.0
PRIMER_MIN_TM=50.0
PRIMER_INTERNAL_MIN_TM=50.0
PRIMER_WT_TM_LT=0.0
PRIMER_INTERNAL_WT_TM_LT=0.0
PRIMER_WT_TM_GT=0.0
PRIMER_INTERNAL_WT_TM_GT=0.0
PRIMER_MIN_BOUND=96.5
PRIMER_OPT_BOUND=97.0
PRIMER_MAX_BOUND=97.5
PRIMER_INTERNAL_MIN_BOUND=96.5
PRIMER_INTERNAL_OPT_BOUND=97.0
PRIMER_INTERNAL_MAX_BOUND=97.5
PRIMER_WT_BOUND_LT=1.0
PRIMER_WT_BOUND_GT=1.0
PRIMER_INTERNAL_WT_BOUND_LT=1.0
PRIMER_INTERNAL_WT_BOUND_GT=1.0
SEQUENCE_ID=Pick primers by bound fraction SantaLucia - SantaLucia
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_LEFT_EXPLAIN=considered 13974, GC content failed 3, low tm 2758, low faction bound 7037, high fraction bound 3502, high any compl 2, high end compl 6, long poly-x seq 44, ok 622
PRIMER_RIGHT_EXPLAIN=considered 13974, GC content failed 3, low tm 2986, low faction bound 7017, high fraction bound 3314, high any compl 1, high end compl 4, long poly-x seq 44, ok 605
PRIMER_INTERNAL_EXPLAIN=considered 14749, GC content failed 3, low tm 3081, low faction bound 7426, high fraction bound 3543, long poly-x seq 44, ok 652
PRIMER_PAIR_EXPLAIN=considered 622, unacceptable product size 614, high end compl 2, ok 6
PRIMER_LEFT_NUM_RETURNED=4
PRIMER_RIGHT_NUM_RETURNED=4
PRIMER_INTERNAL_NUM_RETURNED=4
PRIMER_PAIR_NUM_RETURNED=4
PRIMER_PAIR_0_PENALTY=0.436315
PRIMER_LEFT_0_PENALTY=0.113511
PRIMER_RIGHT_0_PENALTY=0.322804
PRIMER_INTERNAL_0_PENALTY=1.368218
PRIMER_LEFT_0_SEQUENCE=gctggcgcggaagccttaac
PRIMER_RIGHT_0_SEQUENCE=ttatgcgcacccccagcacc
PRIMER_INTERNAL_0_SEQUENCE=gccgctcatttcaccgaagcc
PRIMER_LEFT_0=366,20
PRIMER_RIGHT_0=541,20
PRIMER_INTERNAL_0=483,21
PRIMER_LEFT_0_TM=58.962
PRIMER_RIGHT_0_TM=59.690
PRIMER_INTERNAL_0_TM=59.013
PRIMER_LEFT_0_BOUND=96.886
PRIMER_RIGHT_0_BOUND=97.323
PRIMER_INTERNAL_0_BOUND=97.368
PRIMER_LEFT_0_GC_PERCENT=65.000
PRIMER_RIGHT_0_GC_PERCENT=65.000
PRIMER_INTERNAL_0_GC_PERCENT=61.905
PRIMER_LEFT_0_SELF_ANY=6.00
PRIMER_RIGHT_0_SELF_ANY=6.00
PRIMER_INTERNAL_0_SELF_ANY=3.00
PRIMER_LEFT_0_SELF_END=2.00
PRIMER_RIGHT_0_SELF_END=0.00
PRIMER_INTERNAL_0_SELF_END=1.00
PRIMER_LEFT_0_END_STABILITY=2.0100
PRIMER_RIGHT_0_END_STABILITY=5.0100
PRIMER_PAIR_0_COMPL_ANY=5.00
PRIMER_PAIR_0_COMPL_END=3.00
PRIMER_PAIR_0_PRODUCT_SIZE=176
PRIMER_PAIR_0_PRODUCT_TM=77.7
PRIMER_PAIR_1_PENALTY=0.484172
PRIMER_LEFT_1_PENALTY=0.322804
PRIMER_RIGHT_1_PENALTY=0.161367
PRIMER_INTERNAL_1_PENALTY=2.175315
PRIMER_LEFT_1_SEQUENCE=ggtgctgggggtgcgcataa
PRIMER_RIGHT_1_SEQUENCE=gtgggccacggcttcgatga
PRIMER_INTERNAL_1_SEQUENCE=tcggccacttcacagccaatca
PRIMER_LEFT_1=522,20
PRIMER_RIGHT_1=707,20
PRIMER_INTERNAL_1=661,22
PRIMER_LEFT_1_TM=59.690
PRIMER_RIGHT_1_TM=59.419
PRIMER_INTERNAL_1_TM=58.763
PRIMER_LEFT_1_BOUND=97.323
PRIMER_RIGHT_1_BOUND=97.161
PRIMER_INTERNAL_1_BOUND=97.175
PRIMER_LEFT_1_GC_PERCENT=65.000
PRIMER_RIGHT_1_GC_PERCENT=65.000
PRIMER_INTERNAL_1_GC_PERCENT=54.545
PRIMER_LEFT_1_SELF_ANY=6.00
PRIMER_RIGHT_1_SELF_ANY=7.00
PRIMER_INTERNAL_1_SELF_ANY=4.00
PRIMER_LEFT_1_SELF_END=1.00
PRIMER_RIGHT_1_SELF_END=2.00
PRIMER_INTERNAL_1_SELF_END=1.00
PRIMER_LEFT_1_END_STABILITY=1.9000
PRIMER_RIGHT_1_END_STABILITY=2.9200
PRIMER_PAIR_1_COMPL_ANY=3.00
PRIMER_PAIR_1_COMPL_END=1.00
PRIMER_PAIR_1_PRODUCT_SIZE=186
PRIMER_PAIR_1_PRODUCT_TM=77.2
PRIMER_PAIR_2_PENALTY=0.718795
PRIMER_LEFT_2_PENALTY=0.322804
PRIMER_RIGHT_2_PENALTY=0.395991
PRIMER_INTERNAL_2_PENALTY=2.175315
PRIMER_LEFT_2_SEQUENCE=ggtgctgggggtgcgcataa
PRIMER_RIGHT_2_SEQUENCE=gccacggcttcgatgatggc
PRIMER_INTERNAL_2_SEQUENCE=tcggccacttcacagccaatca
PRIMER_LEFT_2=522,20
PRIMER_RIGHT_2=703,20
PRIMER_INTERNAL_2=661,22
PRIMER_LEFT_2_TM=59.690
PRIMER_RIGHT_2_TM=58.755
PRIMER_INTERNAL_2_TM=58.763
PRIMER_LEFT_2_BOUND=97.323
PRIMER_RIGHT_2_BOUND=96.604
PRIMER_INTERNAL_2_BOUND=97.175
PRIMER_LEFT_2_GC_PERCENT=65.000
PRIMER_RIGHT_2_GC_PERCENT=65.000
PRIMER_INTERNAL_2_GC_PERCENT=54.545
PRIMER_LEFT_2_SELF_ANY=6.00
PRIMER_RIGHT_2_SELF_ANY=4.00
PRIMER_INTERNAL_2_SELF_ANY=4.00
PRIMER_LEFT_2_SELF_END=1.00
PRIMER_RIGHT_2_SELF_END=2.00
PRIMER_INTERNAL_2_SELF_END=1.00
PRIMER_LEFT_2_END_STABILITY=1.9000
PRIMER_RIGHT_2_END_STABILITY=4.4000
PRIMER_PAIR_2_COMPL_ANY=3.00
PRIMER_PAIR_2_COMPL_END=3.00
PRIMER_PAIR_2_PRODUCT_SIZE=182
PRIMER_PAIR_2_PRODUCT_TM=76.9
PRIMER_PAIR_3_PENALTY=1.364650
PRIMER_LEFT_3_PENALTY=1.041846
PRIMER_RIGHT_3_PENALTY=0.322804
PRIMER_INTERNAL_3_PENALTY=1.368218
PRIMER_LEFT_3_SEQUENCE=ggctggcgcggaagcctta
PRIMER_RIGHT_3_SEQUENCE=ttatgcgcacccccagcacc
PRIMER_INTERNAL_3_SEQUENCE=gccgctcatttcaccgaagcc
PRIMER_LEFT_3=365,19
PRIMER_RIGHT_3=541,20
PRIMER_INTERNAL_3=483,21
PRIMER_LEFT_3_TM=59.747
PRIMER_RIGHT_3_TM=59.690
PRIMER_INTERNAL_3_TM=59.013
PRIMER_LEFT_3_BOUND=97.042
PRIMER_RIGHT_3_BOUND=97.323
PRIMER_INTERNAL_3_BOUND=97.368
PRIMER_LEFT_3_GC_PERCENT=68.421
PRIMER_RIGHT_3_GC_PERCENT=65.000
PRIMER_INTERNAL_3_GC_PERCENT=61.905
PRIMER_LEFT_3_SELF_ANY=8.00
PRIMER_RIGHT_3_SELF_ANY=6.00
PRIMER_INTERNAL_3_SELF_ANY=3.00
PRIMER_LEFT_3_SELF_END=3.00
PRIMER_RIGHT_3_SELF_END=0.00
PRIMER_INTERNAL_3_SELF_END=1.00
PRIMER_LEFT_3_END_STABILITY=2.6900
PRIMER_RIGHT_3_END_STABILITY=5.0100
PRIMER_PAIR_3_COMPL_ANY=5.00
PRIMER_PAIR_3_COMPL_END=2.00
PRIMER_PAIR_3_PRODUCT_SIZE=177
PRIMER_PAIR_3_PRODUCT_TM=77.8
=
PRIMER_TM_FORMULA=1
PRIMER_SALT_CORRECTIONS=1
PRIMER_ANNEALING_TEMP=50.0
PRIMER_MIN_TM=40.0
PRIMER_INTERNAL_MIN_TM=40.0
PRIMER_MIN_BOUND=94.5
PRIMER_OPT_BOUND=95.0
PRIMER_MAX_BOUND=95.5
PRIMER_INTERNAL_MIN_BOUND=94.5
PRIMER_INTERNAL_OPT_BOUND=95.0
PRIMER_INTERNAL_MAX_BOUND=95.5
SEQUENCE_ID=Pick primers by bound fraction with low Tm SantaLucia - SantaLucia
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_LEFT_EXPLAIN=considered 13974, GC content failed 3, low tm 15, low faction bound 8808, high fraction bound 4632, high end compl 3, long poly-x seq 44, ok 469
PRIMER_RIGHT_EXPLAIN=considered 13974, GC content failed 3, low tm 25, low faction bound 9020, high fraction bound 4413, high end compl 1, long poly-x seq 44, ok 468
PRIMER_INTERNAL_EXPLAIN=considered 14749, GC content failed 3, low tm 25, low faction bound 9467, high fraction bound 4715, long poly-x seq 44, ok 495
PRIMER_PAIR_EXPLAIN=considered 135, unacceptable product size 122, high end compl 7, ok 6
PRIMER_LEFT_NUM_RETURNED=4
PRIMER_RIGHT_NUM_RETURNED=4
PRIMER_INTERNAL_NUM_RETURNED=4
PRIMER_PAIR_NUM_RETURNED=4
PRIMER_PAIR_0_PENALTY=0.415187
PRIMER_LEFT_0_PENALTY=0.056524
PRIMER_RIGHT_0_PENALTY=0.358663
PRIMER_INTERNAL_0_PENALTY=3.294240
PRIMER_LEFT_0_SEQUENCE=accaccaccacaaccacgga
PRIMER_RIGHT_0_SEQUENCE=cttccgcgccagccatcaat
PRIMER_INTERNAL_0_SEQUENCE=aaccacaacaacaacaacggcaa
PRIMER_LEFT_0=237,20
PRIMER_RIGHT_0=378,20
PRIMER_INTERNAL_0=284,23
PRIMER_LEFT_0_TM=58.114
PRIMER_RIGHT_0_TM=58.040
PRIMER_INTERNAL_0_TM=56.766
PRIMER_LEFT_0_BOUND=95.057
PRIMER_RIGHT_0_BOUND=95.359
PRIMER_INTERNAL_0_BOUND=94.706
PRIMER_LEFT_0_GC_PERCENT=60.000
PRIMER_RIGHT_0_GC_PERCENT=60.000
PRIMER_INTERNAL_0_GC_PERCENT=43.478
PRIMER_LEFT_0_SELF_ANY=2.00
PRIMER_RIGHT_0_SELF_ANY=4.00
PRIMER_INTERNAL_0_SELF_ANY=2.00
PRIMER_LEFT_0_SELF_END=0.00
PRIMER_RIGHT_0_SELF_END=2.00
PRIMER_INTERNAL_0_SELF_END=0.00
PRIMER_LEFT_0_END_STABILITY=4.6900
PRIMER_RIGHT_0_END_STABILITY=2.5700
PRIMER_PAIR_0_COMPL_ANY=4.00
PRIMER_PAIR_0_COMPL_END=1.00
PRIMER_PAIR_0_PRODUCT_SIZE=142
PRIMER_PAIR_0_PRODUCT_TM=74.2
PRIMER_PAIR_1_PENALTY=0.627942
PRIMER_LEFT_1_PENALTY=0.496690
PRIMER_RIGHT_1_PENALTY=0.131252
PRIMER_INTERNAL_1_PENALTY=1.060858
PRIMER_LEFT_1_SEQUENCE=gtgctgggggtgcgcataat
PRIMER_RIGHT_1_SEQUENCE=ggcccattggaccccttcca
PRIMER_INTERNAL_1_SEQUENCE=aagccatcatcgaagccgtgg
PRIMER_LEFT_1=523,20
PRIMER_RIGHT_1=761,20
PRIMER_INTERNAL_1=682,21
PRIMER_LEFT_1_TM=57.663
PRIMER_RIGHT_1_TM=58.296
PRIMER_INTERNAL_1_TM=57.534
PRIMER_LEFT_1_BOUND=94.503
PRIMER_RIGHT_1_BOUND=95.131
PRIMER_INTERNAL_1_BOUND=95.061
PRIMER_LEFT_1_GC_PERCENT=60.000
PRIMER_RIGHT_1_GC_PERCENT=65.000
PRIMER_INTERNAL_1_GC_PERCENT=57.143
PRIMER_LEFT_1_SELF_ANY=6.00
PRIMER_RIGHT_1_SELF_ANY=7.00
PRIMER_INTERNAL_1_SELF_ANY=4.00
PRIMER_LEFT_1_SELF_END=3.00
PRIMER_RIGHT_1_SELF_END=3.00
PRIMER_INTERNAL_1_SELF_END=2.00
PRIMER_LEFT_1_END_STABILITY=1.2800
PRIMER_RIGHT_1_END_STABILITY=3.5300
PRIMER_PAIR_1_COMPL_ANY=5.00
PRIMER_PAIR_1_COMPL_END=1.00
PRIMER_PAIR_1_PRODUCT_SIZE=239
PRIMER_PAIR_1_PRODUCT_TM=78.5
PRIMER_PAIR_2_PENALTY=0.696223
PRIMER_LEFT_2_PENALTY=0.337560
PRIMER_RIGHT_2_PENALTY=0.358663
PRIMER_INTERNAL_2_PENALTY=3.294240
PRIMER_LEFT_2_SEQUENCE=caccaccaccacaaccacgg
PRIMER_RIGHT_2_SEQUENCE=cttccgcgccagccatcaat
PRIMER_INTERNAL_2_SEQUENCE=aaccacaacaacaacaacggcaa
PRIMER_LEFT_2=236,20
PRIMER_RIGHT_2=378,20
PRIMER_INTERNAL_2=284,23
PRIMER_LEFT_2_TM=58.037
PRIMER_RIGHT_2_TM=58.040
PRIMER_INTERNAL_2_TM=56.766
PRIMER_LEFT_2_BOUND=95.338
PRIMER_RIGHT_2_BOUND=95.359
PRIMER_INTERNAL_2_BOUND=94.706
PRIMER_LEFT_2_GC_PERCENT=65.000
PRIMER_RIGHT_2_GC_PERCENT=60.000
PRIMER_INTERNAL_2_GC_PERCENT=43.478
PRIMER_LEFT_2_SELF_ANY=2.00
PRIMER_RIGHT_2_SELF_ANY=4.00
PRIMER_INTERNAL_2_SELF_ANY=2.00
PRIMER_LEFT_2_SELF_END=2.00
PRIMER_RIGHT_2_SELF_END=2.00
PRIMER_INTERNAL_2_SELF_END=0.00
PRIMER_LEFT_2_END_STABILITY=4.9400
PRIMER_RIGHT_2_END_STABILITY=2.5700
PRIMER_PAIR_2_COMPL_ANY=3.00
PRIMER_PAIR_2_COMPL_END=0.00
PRIMER_PAIR_2_PRODUCT_SIZE=143
PRIMER_PAIR_2_PRODUCT_TM=74.3
PRIMER_PAIR_3_PENALTY=0.696223
PRIMER_LEFT_3_PENALTY=0.337560
PRIMER_RIGHT_3_PENALTY=0.358663
PRIMER_INTERNAL_3_PENALTY=3.294240
PRIMER_LEFT_3_SEQUENCE=ccaccaccaccacaaccacg
PRIMER_RIGHT_3_SEQUENCE=cttccgcgccagccatcaat
PRIMER_INTERNAL_3_SEQUENCE=aaccacaacaacaacaacggcaa
PRIMER_LEFT_3=235,20
PRIMER_RIGHT_3=378,20
PRIMER_INTERNAL_3=284,23
PRIMER_LEFT_3_TM=58.037
PRIMER_RIGHT_3_TM=58.040
PRIMER_INTERNAL_3_TM=56.766
PRIMER_LEFT_3_BOUND=95.338
PRIMER_RIGHT_3_BOUND=95.359
PRIMER_INTERNAL_3_BOUND=94.706
PRIMER_LEFT_3_GC_PERCENT=65.000
PRIMER_RIGHT_3_GC_PERCENT=60.000
PRIMER_INTERNAL_3_GC_PERCENT=43.478
PRIMER_LEFT_3_SELF_ANY=2.00
PRIMER_RIGHT_3_SELF_ANY=4.00
PRIMER_INTERNAL_3_SELF_ANY=2.00
PRIMER_LEFT_3_SELF_END=2.00
PRIMER_RIGHT_3_SELF_END=2.00
PRIMER_INTERNAL_3_SELF_END=0.00
PRIMER_LEFT_3_END_STABILITY=4.9400
PRIMER_RIGHT_3_END_STABILITY=2.5700
PRIMER_PAIR_3_COMPL_ANY=2.00
PRIMER_PAIR_3_COMPL_END=0.00
PRIMER_PAIR_3_PRODUCT_SIZE=144
PRIMER_PAIR_3_PRODUCT_TM=74.5
=
