MARKER_NAME=boundary_test_00
PRIMER_MAX_POLY_X=0
PRIMER_INTERNAL_OLIGO_MAX_POLY_X=0
PRIMER_EXPLAIN_FLAG=1
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_PRODUCT_SIZE_RANGE=50-100
PRIMER_MIN_TM=0.0
PRIMER_MAX_TM=100.0
PRIMER_MIN_SIZE=25
PRIMER_MAX_SIZE=25
PRIMER_INTERNAL_OLIGO_MIN_SIZE=25
PRIMER_INTERNAL_OLIGO_MAX_SIZE=25
PRIMER_INTERNAL_OLIGO_OPT_SIZE=25
PRIMER_OPT_SIZE=25
PRIMER_MAX_GC=100
PRIMER_MIN_GC=0
PRIMER_SELF_END=100
PRIMER_SELF_ANY=100
PRIMER_INTERNAL_OLIGO_SELF_ANY=100
PRIMER_INTERNAL_OLIGO_SELF_END=100 
PRIMER_INTERNAL_OLIGO_MAX_TM=100
PRIMER_INTERNAL_OLIGO_MIN_TM=30
PRIMER_INTERNAL_OLIGO_MAX_GC=100
PRIMER_INTERNAL_OLIGO_MIN_GC=0
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_LEFT_EXPLAIN=considered 35, too many Ns 33, ok 2
PRIMER_RIGHT_EXPLAIN=considered 35, too many Ns 33, ok 2
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 60, too many Ns 57, ok 3
PRIMER_PAIR_EXPLAIN=considered 4, unacceptable product size 1, no internal oligo 2, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=11.495378
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=ACGTACGTACGTTGCAGTCGACGCA
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=30,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=71.495
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=56.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=8.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_01
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNCACGTCATCTCATCAAATTTGATTGNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_PRODUCT_SIZE_RANGE=55-100
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_LEFT_EXPLAIN=considered 30, too many Ns 28, ok 2
PRIMER_RIGHT_EXPLAIN=considered 30, too many Ns 29, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 60, too many Ns 57, ok 3
PRIMER_PAIR_EXPLAIN=considered 2, unacceptable product size 1, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=2.731673
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=29,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=62.732
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=36.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=10.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_02
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNCACGTCATCTCATCAAATTTGATTGNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_PRODUCT_SIZE_RANGE=80-80
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_LEFT_EXPLAIN=considered 5, too many Ns 4, ok 1
PRIMER_RIGHT_EXPLAIN=considered 5, too many Ns 4, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 60, too many Ns 57, ok 3
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=2.731673
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=29,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=62.732
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=36.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=10.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_05
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNCACGTCATCTCATCAAATTTGATTGNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_PRODUCT_SIZE_RANGE=80-80
PRIMER_MIN_SIZE=24
PRIMER_MAX_SIZE=24
PRIMER_OPT_SIZE=24
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_LEFT_EXPLAIN=considered 5, too many Ns 3, ok 2
PRIMER_RIGHT_EXPLAIN=considered 5, too many Ns 3, ok 2
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 60, too many Ns 57, ok 3
PRIMER_PAIR_EXPLAIN=considered 4, unacceptable product size 3, ok 1
PRIMER_PAIR_PENALTY=0.3310
PRIMER_LEFT_PENALTY=0.243965
PRIMER_RIGHT_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_PENALTY=2.731673
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATT
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_LEFT=2,24
PRIMER_RIGHT=81,24
PRIMER_INTERNAL_OLIGO=29,25
PRIMER_LEFT_TM=60.244
PRIMER_RIGHT_TM=59.913
PRIMER_INTERNAL_OLIGO_TM=62.732
PRIMER_LEFT_GC_PERCENT=33.333
PRIMER_RIGHT_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_GC_PERCENT=36.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_SELF_END=10.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=2.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_11
PRIMER_NUM_NS_ACCEPTED=1
SEQUENCE=NNCACGTCATCTNATCAAATTTGATTGNNCACGTCATCTNATCAAATTTGATTGNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_LEFT_EXPLAIN=considered 5, too many Ns 3, ok 2
PRIMER_RIGHT_EXPLAIN=considered 5, too many Ns 1, ok 4
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 60, too many Ns 59, ok 1
PRIMER_PAIR_EXPLAIN=considered 8, unacceptable product size 6, no internal oligo 2, ok 0
=
MARKER_NAME=boundary_test_11
PRIMER_NUM_NS_ACCEPTED=2
PRIMER_INTERNAL_OLIGO_NUM_NS=2
SEQUENCE=NNCACGTCATCNNATCAAATTTGATTGNNCACGTCATCNNATCAAATTTGATTGNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_LEFT_EXPLAIN=considered 5, too many Ns 3, ok 2
PRIMER_RIGHT_EXPLAIN=considered 5, ok 5
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 60, too many Ns 52, ok 8
PRIMER_PAIR_EXPLAIN=considered 10, unacceptable product size 8, ok 2
PRIMER_PAIR_PENALTY=0.8938
PRIMER_LEFT_PENALTY=0.806819
PRIMER_RIGHT_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_PENALTY=3.221174
PRIMER_LEFT_SEQUENCE=CACGTCATCNNATCAAATTTGATT
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_SEQUENCE=CACGTCATCNNATCAAATTTGATTG
PRIMER_LEFT=2,24
PRIMER_RIGHT=81,24
PRIMER_INTERNAL_OLIGO=29,25
PRIMER_LEFT_TM=60.807
PRIMER_RIGHT_TM=59.913
PRIMER_INTERNAL_OLIGO_TM=63.221
PRIMER_LEFT_GC_PERCENT=31.818
PRIMER_RIGHT_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_GC_PERCENT=34.783
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=11.50
PRIMER_RIGHT_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_SELF_END=11.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=6.00
PRIMER_PAIR_COMPL_END=2.00
PRIMER_PRODUCT_SIZE=80
PRIMER_PAIR_PENALTY_1=2.5470
PRIMER_LEFT_1_PENALTY=0.806819
PRIMER_RIGHT_1_PENALTY=1.740140
PRIMER_INTERNAL_OLIGO_1_PENALTY=3.221174
PRIMER_LEFT_1_SEQUENCE=ACGTCATCNNATCAAATTTGATTG
PRIMER_RIGHT_1_SEQUENCE=NGAATTGTTTGGAAATGACAGTGA
PRIMER_INTERNAL_OLIGO_1_SEQUENCE=CACGTCATCNNATCAAATTTGATTG
PRIMER_LEFT_1=3,24
PRIMER_RIGHT_1=82,24
PRIMER_INTERNAL_OLIGO_1=29,25
PRIMER_LEFT_1_TM=60.807
PRIMER_RIGHT_1_TM=61.740
PRIMER_INTERNAL_OLIGO_1_TM=63.221
PRIMER_LEFT_1_GC_PERCENT=31.818
PRIMER_RIGHT_1_GC_PERCENT=34.783
PRIMER_INTERNAL_OLIGO_1_GC_PERCENT=34.783
PRIMER_LEFT_1_SELF_ANY=12.00
PRIMER_RIGHT_1_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_1_SELF_ANY=12.00
PRIMER_LEFT_1_SELF_END=11.00
PRIMER_RIGHT_1_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_1_SELF_END=11.00
PRIMER_LEFT_1_END_STABILITY=6.9000
PRIMER_RIGHT_1_END_STABILITY=6.4000
PRIMER_PAIR_1_COMPL_ANY=5.00
PRIMER_PAIR_1_COMPL_END=2.00
PRIMER_PRODUCT_SIZE_1=80
=
MARKER_NAME=boundary_test_12
PRIMER_NUM_NS_ACCEPTED=2
SEQUENCE=NNCACGTCATCNNATCAAATTTGATTGNNCACGTCATCNNATCAAATTTGATTGNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_LEFT_EXPLAIN=considered 5, too many Ns 3, ok 2
PRIMER_RIGHT_EXPLAIN=considered 5, ok 5
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 60, too many Ns 52, ok 8
PRIMER_PAIR_EXPLAIN=considered 10, unacceptable product size 8, ok 2
PRIMER_PAIR_PENALTY=0.8938
PRIMER_LEFT_PENALTY=0.806819
PRIMER_RIGHT_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_PENALTY=3.221174
PRIMER_LEFT_SEQUENCE=CACGTCATCNNATCAAATTTGATT
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_SEQUENCE=CACGTCATCNNATCAAATTTGATTG
PRIMER_LEFT=2,24
PRIMER_RIGHT=81,24
PRIMER_INTERNAL_OLIGO=29,25
PRIMER_LEFT_TM=60.807
PRIMER_RIGHT_TM=59.913
PRIMER_INTERNAL_OLIGO_TM=63.221
PRIMER_LEFT_GC_PERCENT=31.818
PRIMER_RIGHT_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_GC_PERCENT=34.783
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=11.50
PRIMER_RIGHT_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_SELF_END=11.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=6.00
PRIMER_PAIR_COMPL_END=2.00
PRIMER_PRODUCT_SIZE=80
PRIMER_PAIR_PENALTY_1=2.5470
PRIMER_LEFT_1_PENALTY=0.806819
PRIMER_RIGHT_1_PENALTY=1.740140
PRIMER_INTERNAL_OLIGO_1_PENALTY=3.221174
PRIMER_LEFT_1_SEQUENCE=ACGTCATCNNATCAAATTTGATTG
PRIMER_RIGHT_1_SEQUENCE=NGAATTGTTTGGAAATGACAGTGA
PRIMER_INTERNAL_OLIGO_1_SEQUENCE=CACGTCATCNNATCAAATTTGATTG
PRIMER_LEFT_1=3,24
PRIMER_RIGHT_1=82,24
PRIMER_INTERNAL_OLIGO_1=29,25
PRIMER_LEFT_1_TM=60.807
PRIMER_RIGHT_1_TM=61.740
PRIMER_INTERNAL_OLIGO_1_TM=63.221
PRIMER_LEFT_1_GC_PERCENT=31.818
PRIMER_RIGHT_1_GC_PERCENT=34.783
PRIMER_INTERNAL_OLIGO_1_GC_PERCENT=34.783
PRIMER_LEFT_1_SELF_ANY=12.00
PRIMER_RIGHT_1_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_1_SELF_ANY=12.00
PRIMER_LEFT_1_SELF_END=11.00
PRIMER_RIGHT_1_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_1_SELF_END=11.00
PRIMER_LEFT_1_END_STABILITY=6.9000
PRIMER_RIGHT_1_END_STABILITY=6.4000
PRIMER_PAIR_1_COMPL_ANY=5.00
PRIMER_PAIR_1_COMPL_END=2.00
PRIMER_PRODUCT_SIZE_1=80
=
MARKER_NAME=boundary_test_13
PRIMER_NUM_NS_ACCEPTED=0
PRIMER_MIN_GC=0
SEQUENCE=NNAAAAAAAAAAAAAAAAAAAAAAAAANNTTTTTTTTTTTTTTTTTTTTTTTTTNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_LEFT_EXPLAIN=considered 5, too many Ns 3, ok 2
PRIMER_RIGHT_EXPLAIN=considered 5, too many Ns 3, ok 2
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 60, too many Ns 23, ok 37
PRIMER_PAIR_EXPLAIN=considered 4, unacceptable product size 3, ok 1
PRIMER_PAIR_PENALTY=5.3942
PRIMER_LEFT_PENALTY=5.307199
PRIMER_RIGHT_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_PENALTY=3.800256
PRIMER_LEFT_SEQUENCE=AAAAAAAAAAAAAAAAAAAAAAAA
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_SEQUENCE=TTTTTTTTTTTTTTTTTTTTTTTNN
PRIMER_LEFT=2,24
PRIMER_RIGHT=81,24
PRIMER_INTERNAL_OLIGO=31,25
PRIMER_LEFT_TM=54.693
PRIMER_RIGHT_TM=59.913
PRIMER_INTERNAL_OLIGO_TM=56.200
PRIMER_LEFT_GC_PERCENT=0.000
PRIMER_RIGHT_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_GC_PERCENT=0.000
PRIMER_LEFT_SELF_ANY=0.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=0.00
PRIMER_LEFT_SELF_END=0.00
PRIMER_RIGHT_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_SELF_END=0.00
PRIMER_LEFT_END_STABILITY=7.6000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=4.00
PRIMER_PAIR_COMPL_END=0.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_14
PRIMER_NUM_NS_ACCEPTED=0
PRIMER_MIN_GC=.0001
SEQUENCE=NNAAAAAAAAAAAAAAAAACAAAAAAATTTTTTTTTTTTTTTTTTTTTTTTTNNNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_LEFT_EXPLAIN=considered 5, too many Ns 2, ok 3
PRIMER_RIGHT_EXPLAIN=considered 5, too many Ns 3, ok 2
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 60, too many Ns 25, ok 35
PRIMER_PAIR_EXPLAIN=considered 6, unacceptable product size 5, ok 1
PRIMER_PAIR_PENALTY=4.8436
PRIMER_LEFT_PENALTY=4.756523
PRIMER_RIGHT_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_PENALTY=3.800256
PRIMER_LEFT_SEQUENCE=AAAAAAAAAAAAAAAAACAAAAAA
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_SEQUENCE=TTTTTTTTTTTTTTTTTTTTTTTNN
PRIMER_LEFT=2,24
PRIMER_RIGHT=81,24
PRIMER_INTERNAL_OLIGO=29,25
PRIMER_LEFT_TM=55.243
PRIMER_RIGHT_TM=59.913
PRIMER_INTERNAL_OLIGO_TM=56.200
PRIMER_LEFT_GC_PERCENT=4.167
PRIMER_RIGHT_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_GC_PERCENT=0.000
PRIMER_LEFT_SELF_ANY=0.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=0.00
PRIMER_LEFT_SELF_END=0.00
PRIMER_RIGHT_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_SELF_END=0.00
PRIMER_LEFT_END_STABILITY=7.6000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=6.00
PRIMER_PAIR_COMPL_END=0.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_15
PRIMER_NUM_NS_ACCEPTED=0
PRIMER_MIN_GC=20
PRIMER_MAX_GC=100
SEQUENCE=NNGGGGGGGGGGGGGGGGGGGGGGGGGNNNNNTTTTTTTTTTTTTTTTTTTTTTTTTACTCACTGTCATTTCCAAACAATTCNN
PRIMER_LEFT_EXPLAIN=considered 5, too many Ns 3, ok 2
PRIMER_RIGHT_EXPLAIN=considered 5, too many Ns 2, ok 3
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 60, too many Ns 25, ok 35
PRIMER_PAIR_EXPLAIN=considered 6, unacceptable product size 5, ok 1
PRIMER_PAIR_PENALTY=29.3957
PRIMER_LEFT_PENALTY=29.308664
PRIMER_RIGHT_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_PENALTY=4.161950
PRIMER_LEFT_SEQUENCE=GGGGGGGGGGGGGGGGGGGGGGGG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_SEQUENCE=TTTTTTTTTTTTTTTTTTTTTTTTT
PRIMER_LEFT=2,24
PRIMER_RIGHT=81,24
PRIMER_INTERNAL_OLIGO=32,25
PRIMER_LEFT_TM=89.309
PRIMER_RIGHT_TM=59.913
PRIMER_INTERNAL_OLIGO_TM=55.838
PRIMER_LEFT_GC_PERCENT=100.000
PRIMER_RIGHT_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_GC_PERCENT=0.000
PRIMER_LEFT_SELF_ANY=0.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=0.00
PRIMER_LEFT_SELF_END=0.00
PRIMER_RIGHT_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_SELF_END=0.00
PRIMER_LEFT_END_STABILITY=12.4000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=1.00
PRIMER_PAIR_COMPL_END=0.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_16
PRIMER_NUM_NS_ACCEPTED=0
PRIMER_PRODUCT_SIZE_RANGE=50-200
PRIMER_INTERNAL_OLIGO_NUM_NS=5
SEQUENCE=NNGGGGGGGGGGGGGGGGGGGGGGGGGNNNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_LEFT_EXPLAIN=considered 10, too many Ns 8, ok 2
PRIMER_RIGHT_EXPLAIN=considered 10, too many Ns 8, ok 2
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 35, ok 35
PRIMER_PAIR_EXPLAIN=considered 4, no internal oligo 4, ok 0
=
MARKER_NAME=boundary_test_17
PRIMER_NUM_NS_ACCEPTED=0
PRIMER_MIN_GC=0
PRIMER_MAX_GC=100
PRIMER_INTERNAL_OLIGO_MIN_GC=20
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNTTTTTTTTTTTTTTTTTTTTTTTTGNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_LEFT_EXPLAIN=considered 35, too many Ns 31, ok 4
PRIMER_RIGHT_EXPLAIN=considered 35, too many Ns 31, ok 4
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 60, GC content failed 29, ok 31
PRIMER_PAIR_EXPLAIN=considered 16, unacceptable product size 4, no internal oligo 12, ok 0
=
MARKER_NAME=boundary_test_18
PRIMER_NUM_NS_ACCEPTED=0
PRIMER_MIN_GC=0
PRIMER_MAX_GC=100
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=10,50
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNACCGCTATTGGGCCATTTGCCCATTGCATGNNNNNNNNNNNNNNNNNNNNNNNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_LEFT_EXPLAIN=considered 62, too many Ns 53, ok 9
PRIMER_RIGHT_EXPLAIN=considered 62, too many Ns 53, ok 9
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 87, too many Ns 39, overlap excluded region 40, ok 8
PRIMER_PAIR_EXPLAIN=considered 81, unacceptable product size 49, no internal oligo 32, ok 0
=
MARKER_NAME=boundary_test_19
PRIMER_NUM_NS_ACCEPTED=0
PRIMER_MIN_GC=0
PRIMER_MAX_GC=100
PRIMER_INTERNAL_OLIGO_NUM_NS=0
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=10,50
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNAAGCTAAGCTTGCCCGTATCCCCTGGATNNNCCGTTAGGTGCTAGCCGTGTAGATTNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_LEFT_EXPLAIN=considered 66, too many Ns 57, ok 9
PRIMER_RIGHT_EXPLAIN=considered 66, too many Ns 57, ok 9
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 91, too many Ns 84, overlap excluded region 5, ok 2
PRIMER_PAIR_EXPLAIN=considered 18, unacceptable product size 3, no internal oligo 8, ok 7
PRIMER_PAIR_PENALTY=0.3310
PRIMER_LEFT_PENALTY=0.243965
PRIMER_RIGHT_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_PENALTY=5.578209
PRIMER_LEFT_SEQUENCE=ACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_SEQUENCE=CCGTTAGGTGCTAGCCGTGTAGATT
PRIMER_LEFT=3,24
PRIMER_RIGHT=112,24
PRIMER_INTERNAL_OLIGO=60,25
PRIMER_LEFT_TM=60.244
PRIMER_RIGHT_TM=59.913
PRIMER_INTERNAL_OLIGO_TM=65.578
PRIMER_LEFT_GC_PERCENT=33.333
PRIMER_RIGHT_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_GC_PERCENT=52.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=6.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_SELF_END=1.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=6.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=110
PRIMER_PAIR_PENALTY_1=0.3310
PRIMER_LEFT_1_PENALTY=0.243965
PRIMER_RIGHT_1_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_1_PENALTY=5.578209
PRIMER_LEFT_1_SEQUENCE=CACGTCATCTCATCAAATTTGATT
PRIMER_RIGHT_1_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_1_SEQUENCE=CCGTTAGGTGCTAGCCGTGTAGATT
PRIMER_LEFT_1=2,24
PRIMER_RIGHT_1=112,24
PRIMER_INTERNAL_OLIGO_1=60,25
PRIMER_LEFT_1_TM=60.244
PRIMER_RIGHT_1_TM=59.913
PRIMER_INTERNAL_OLIGO_1_TM=65.578
PRIMER_LEFT_1_GC_PERCENT=33.333
PRIMER_RIGHT_1_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_1_GC_PERCENT=52.000
PRIMER_LEFT_1_SELF_ANY=12.00
PRIMER_RIGHT_1_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_1_SELF_ANY=6.00
PRIMER_LEFT_1_SELF_END=10.00
PRIMER_RIGHT_1_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_1_SELF_END=1.00
PRIMER_LEFT_1_END_STABILITY=6.9000
PRIMER_RIGHT_1_END_STABILITY=6.4000
PRIMER_PAIR_1_COMPL_ANY=7.00
PRIMER_PAIR_1_COMPL_END=2.00
PRIMER_PRODUCT_SIZE_1=111
PRIMER_PAIR_PENALTY_2=1.2615
PRIMER_LEFT_2_PENALTY=0.243965
PRIMER_RIGHT_2_PENALTY=1.017510
PRIMER_INTERNAL_OLIGO_2_PENALTY=5.578209
PRIMER_LEFT_2_SEQUENCE=ACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_2_SEQUENCE=AATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_2_SEQUENCE=CCGTTAGGTGCTAGCCGTGTAGATT
PRIMER_LEFT_2=3,24
PRIMER_RIGHT_2=111,24
PRIMER_INTERNAL_OLIGO_2=60,25
PRIMER_LEFT_2_TM=60.244
PRIMER_RIGHT_2_TM=58.982
PRIMER_INTERNAL_OLIGO_2_TM=65.578
PRIMER_LEFT_2_GC_PERCENT=33.333
PRIMER_RIGHT_2_GC_PERCENT=33.333
PRIMER_INTERNAL_OLIGO_2_GC_PERCENT=52.000
PRIMER_LEFT_2_SELF_ANY=12.00
PRIMER_RIGHT_2_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_2_SELF_ANY=6.00
PRIMER_LEFT_2_SELF_END=10.00
PRIMER_RIGHT_2_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_2_SELF_END=1.00
PRIMER_LEFT_2_END_STABILITY=6.9000
PRIMER_RIGHT_2_END_STABILITY=6.4000
PRIMER_PAIR_2_COMPL_ANY=6.00
PRIMER_PAIR_2_COMPL_END=1.00
PRIMER_PRODUCT_SIZE_2=109
PRIMER_PAIR_PENALTY_3=1.2615
PRIMER_LEFT_3_PENALTY=0.243965
PRIMER_RIGHT_3_PENALTY=1.017510
PRIMER_INTERNAL_OLIGO_3_PENALTY=5.578209
PRIMER_LEFT_3_SEQUENCE=CACGTCATCTCATCAAATTTGATT
PRIMER_RIGHT_3_SEQUENCE=AATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_3_SEQUENCE=CCGTTAGGTGCTAGCCGTGTAGATT
PRIMER_LEFT_3=2,24
PRIMER_RIGHT_3=111,24
PRIMER_INTERNAL_OLIGO_3=60,25
PRIMER_LEFT_3_TM=60.244
PRIMER_RIGHT_3_TM=58.982
PRIMER_INTERNAL_OLIGO_3_TM=65.578
PRIMER_LEFT_3_GC_PERCENT=33.333
PRIMER_RIGHT_3_GC_PERCENT=33.333
PRIMER_INTERNAL_OLIGO_3_GC_PERCENT=52.000
PRIMER_LEFT_3_SELF_ANY=12.00
PRIMER_RIGHT_3_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_3_SELF_ANY=6.00
PRIMER_LEFT_3_SELF_END=10.00
PRIMER_RIGHT_3_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_3_SELF_END=1.00
PRIMER_LEFT_3_END_STABILITY=6.9000
PRIMER_RIGHT_3_END_STABILITY=6.4000
PRIMER_PAIR_3_COMPL_ANY=7.00
PRIMER_PAIR_3_COMPL_END=2.00
PRIMER_PRODUCT_SIZE_3=110
PRIMER_PAIR_PENALTY_4=8.0059
PRIMER_LEFT_4_PENALTY=7.918829
PRIMER_RIGHT_4_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_4_PENALTY=5.578209
PRIMER_LEFT_4_SEQUENCE=AAGCTAAGCTTGCCCGTATCCCCT
PRIMER_RIGHT_4_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_4_SEQUENCE=CCGTTAGGTGCTAGCCGTGTAGATT
PRIMER_LEFT_4=29,24
PRIMER_RIGHT_4=112,24
PRIMER_INTERNAL_OLIGO_4=60,25
PRIMER_LEFT_4_TM=67.919
PRIMER_RIGHT_4_TM=59.913
PRIMER_INTERNAL_OLIGO_4_TM=65.578
PRIMER_LEFT_4_GC_PERCENT=54.167
PRIMER_RIGHT_4_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_4_GC_PERCENT=52.000
PRIMER_LEFT_4_SELF_ANY=9.00
PRIMER_RIGHT_4_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_4_SELF_ANY=6.00
PRIMER_LEFT_4_SELF_END=0.00
PRIMER_RIGHT_4_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_4_SELF_END=1.00
PRIMER_LEFT_4_END_STABILITY=10.9000
PRIMER_RIGHT_4_END_STABILITY=6.4000
PRIMER_PAIR_4_COMPL_ANY=3.00
PRIMER_PAIR_4_COMPL_END=2.00
PRIMER_PRODUCT_SIZE_4=84
=
MARKER_NAME=boundary_test_20
PRIMER_NUM_NS_ACCEPTED=0
PRIMER_MIN_GC=0
PRIMER_MAX_GC=100
PRIMER_PRODUCT_SIZE_RANGE=100-300
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=5,50
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=60,15
INCLUDED_REGION=2,250
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNAAGCTAAGCTGTCGATGCGGTGGAGACGATACGGTAAGCTAGCTAACTGGTCAAAGCTAGCGGTAGCTTAGCTNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNACTCACTGTCATTTCCAAACAATTCATCTCTCTCTCATATATATATATATATATATATATATAT
PRIMER_LEFT_EXPLAIN=considered 151, too many Ns 99, ok 52
PRIMER_RIGHT_EXPLAIN=considered 151, too many Ns 148, ok 3
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 226, too many Ns 175, overlap excluded region 47, ok 4
PRIMER_PAIR_EXPLAIN=considered 23, no internal oligo 15, ok 8
PRIMER_PAIR_PENALTY=0.3310
PRIMER_LEFT_PENALTY=0.243965
PRIMER_RIGHT_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_PENALTY=3.811648
PRIMER_LEFT_SEQUENCE=ACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_SEQUENCE=CTGGTCAAAGCTAGCGGTAGCTTAG
PRIMER_LEFT=3,24
PRIMER_RIGHT=251,24
PRIMER_INTERNAL_OLIGO=75,25
PRIMER_LEFT_TM=60.244
PRIMER_RIGHT_TM=59.913
PRIMER_INTERNAL_OLIGO_TM=63.812
PRIMER_LEFT_GC_PERCENT=33.333
PRIMER_RIGHT_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_GC_PERCENT=52.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_SELF_END=12.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=6.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=249
PRIMER_PAIR_PENALTY_1=0.3310
PRIMER_LEFT_1_PENALTY=0.243965
PRIMER_RIGHT_1_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_1_PENALTY=3.811648
PRIMER_LEFT_1_SEQUENCE=CACGTCATCTCATCAAATTTGATT
PRIMER_RIGHT_1_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_1_SEQUENCE=CTGGTCAAAGCTAGCGGTAGCTTAG
PRIMER_LEFT_1=2,24
PRIMER_RIGHT_1=251,24
PRIMER_INTERNAL_OLIGO_1=75,25
PRIMER_LEFT_1_TM=60.244
PRIMER_RIGHT_1_TM=59.913
PRIMER_INTERNAL_OLIGO_1_TM=63.812
PRIMER_LEFT_1_GC_PERCENT=33.333
PRIMER_RIGHT_1_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_1_GC_PERCENT=52.000
PRIMER_LEFT_1_SELF_ANY=12.00
PRIMER_RIGHT_1_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_1_SELF_ANY=12.00
PRIMER_LEFT_1_SELF_END=10.00
PRIMER_RIGHT_1_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_1_SELF_END=12.00
PRIMER_LEFT_1_END_STABILITY=6.9000
PRIMER_RIGHT_1_END_STABILITY=6.4000
PRIMER_PAIR_1_COMPL_ANY=7.00
PRIMER_PAIR_1_COMPL_END=2.00
PRIMER_PRODUCT_SIZE_1=250
PRIMER_PAIR_PENALTY_2=0.3864
PRIMER_LEFT_2_PENALTY=0.299404
PRIMER_RIGHT_2_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_2_PENALTY=3.811648
PRIMER_LEFT_2_SEQUENCE=GGAGACGATACGGTAAGCTAGCTA
PRIMER_RIGHT_2_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_2_SEQUENCE=CTGGTCAAAGCTAGCGGTAGCTTAG
PRIMER_LEFT_2=50,24
PRIMER_RIGHT_2=251,24
PRIMER_INTERNAL_OLIGO_2=75,25
PRIMER_LEFT_2_TM=60.299
PRIMER_RIGHT_2_TM=59.913
PRIMER_INTERNAL_OLIGO_2_TM=63.812
PRIMER_LEFT_2_GC_PERCENT=50.000
PRIMER_RIGHT_2_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_2_GC_PERCENT=52.000
PRIMER_LEFT_2_SELF_ANY=8.00
PRIMER_RIGHT_2_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_2_SELF_ANY=12.00
PRIMER_LEFT_2_SELF_END=6.00
PRIMER_RIGHT_2_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_2_SELF_END=12.00
PRIMER_LEFT_2_END_STABILITY=7.2000
PRIMER_RIGHT_2_END_STABILITY=6.4000
PRIMER_PAIR_2_COMPL_ANY=3.00
PRIMER_PAIR_2_COMPL_END=1.00
PRIMER_PRODUCT_SIZE_2=202
PRIMER_PAIR_PENALTY_3=1.2615
PRIMER_LEFT_3_PENALTY=0.243965
PRIMER_RIGHT_3_PENALTY=1.017510
PRIMER_INTERNAL_OLIGO_3_PENALTY=3.811648
PRIMER_LEFT_3_SEQUENCE=ACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_3_SEQUENCE=AATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_3_SEQUENCE=CTGGTCAAAGCTAGCGGTAGCTTAG
PRIMER_LEFT_3=3,24
PRIMER_RIGHT_3=250,24
PRIMER_INTERNAL_OLIGO_3=75,25
PRIMER_LEFT_3_TM=60.244
PRIMER_RIGHT_3_TM=58.982
PRIMER_INTERNAL_OLIGO_3_TM=63.812
PRIMER_LEFT_3_GC_PERCENT=33.333
PRIMER_RIGHT_3_GC_PERCENT=33.333
PRIMER_INTERNAL_OLIGO_3_GC_PERCENT=52.000
PRIMER_LEFT_3_SELF_ANY=12.00
PRIMER_RIGHT_3_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_3_SELF_ANY=12.00
PRIMER_LEFT_3_SELF_END=10.00
PRIMER_RIGHT_3_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_3_SELF_END=12.00
PRIMER_LEFT_3_END_STABILITY=6.9000
PRIMER_RIGHT_3_END_STABILITY=6.4000
PRIMER_PAIR_3_COMPL_ANY=6.00
PRIMER_PAIR_3_COMPL_END=1.00
PRIMER_PRODUCT_SIZE_3=248
PRIMER_PAIR_PENALTY_4=1.2615
PRIMER_LEFT_4_PENALTY=0.243965
PRIMER_RIGHT_4_PENALTY=1.017510
PRIMER_INTERNAL_OLIGO_4_PENALTY=3.811648
PRIMER_LEFT_4_SEQUENCE=CACGTCATCTCATCAAATTTGATT
PRIMER_RIGHT_4_SEQUENCE=AATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_4_SEQUENCE=CTGGTCAAAGCTAGCGGTAGCTTAG
PRIMER_LEFT_4=2,24
PRIMER_RIGHT_4=250,24
PRIMER_INTERNAL_OLIGO_4=75,25
PRIMER_LEFT_4_TM=60.244
PRIMER_RIGHT_4_TM=58.982
PRIMER_INTERNAL_OLIGO_4_TM=63.812
PRIMER_LEFT_4_GC_PERCENT=33.333
PRIMER_RIGHT_4_GC_PERCENT=33.333
PRIMER_INTERNAL_OLIGO_4_GC_PERCENT=52.000
PRIMER_LEFT_4_SELF_ANY=12.00
PRIMER_RIGHT_4_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_4_SELF_ANY=12.00
PRIMER_LEFT_4_SELF_END=10.00
PRIMER_RIGHT_4_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_4_SELF_END=12.00
PRIMER_LEFT_4_END_STABILITY=6.9000
PRIMER_RIGHT_4_END_STABILITY=6.4000
PRIMER_PAIR_4_COMPL_ANY=7.00
PRIMER_PAIR_4_COMPL_END=2.00
PRIMER_PRODUCT_SIZE_4=249
=
MARKER_NAME=boundary_test_21
PRIMER_NUM_NS_ACCEPTED=0
PRIMER_MIN_GC=0
PRIMER_MAX_GC=100
PRIMER_INTERNAL_OLIGO_MIN_GC=0
PRIMER_INTERNAL_OLIGO_MIN_TM=55.7
PRIMER_INTERNAL_OLIGO_MAX_TM=55.9
PRIMER_INTERNAL_OLIGO_OPT_TM=55.8
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGAAAAAAAAAAAAAAAAAAAAAAAAAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGNNNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_LEFT_EXPLAIN=considered 77, too many Ns 27, ok 50
PRIMER_RIGHT_EXPLAIN=considered 77, too many Ns 28, ok 49
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 152, too many Ns 57, low tm 1, high tm 93, ok 1
PRIMER_PAIR_EXPLAIN=considered 323, unacceptable product size 223, no internal oligo 87, ok 13
PRIMER_PAIR_PENALTY=0.3310
PRIMER_LEFT_PENALTY=0.243965
PRIMER_RIGHT_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_PENALTY=0.038050
PRIMER_LEFT_SEQUENCE=ACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_SEQUENCE=AAAAAAAAAAAAAAAAAAAAAAAAA
PRIMER_LEFT=3,24
PRIMER_RIGHT=173,24
PRIMER_INTERNAL_OLIGO=61,25
PRIMER_LEFT_TM=60.244
PRIMER_RIGHT_TM=59.913
PRIMER_INTERNAL_OLIGO_TM=55.838
PRIMER_LEFT_GC_PERCENT=33.333
PRIMER_RIGHT_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_GC_PERCENT=0.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=0.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_SELF_END=0.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=6.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=171
PRIMER_PAIR_PENALTY_1=0.3310
PRIMER_LEFT_1_PENALTY=0.243965
PRIMER_RIGHT_1_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_1_PENALTY=0.038050
PRIMER_LEFT_1_SEQUENCE=CACGTCATCTCATCAAATTTGATT
PRIMER_RIGHT_1_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_1_SEQUENCE=AAAAAAAAAAAAAAAAAAAAAAAAA
PRIMER_LEFT_1=2,24
PRIMER_RIGHT_1=173,24
PRIMER_INTERNAL_OLIGO_1=61,25
PRIMER_LEFT_1_TM=60.244
PRIMER_RIGHT_1_TM=59.913
PRIMER_INTERNAL_OLIGO_1_TM=55.838
PRIMER_LEFT_1_GC_PERCENT=33.333
PRIMER_RIGHT_1_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_1_GC_PERCENT=0.000
PRIMER_LEFT_1_SELF_ANY=12.00
PRIMER_RIGHT_1_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_1_SELF_ANY=0.00
PRIMER_LEFT_1_SELF_END=10.00
PRIMER_RIGHT_1_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_1_SELF_END=0.00
PRIMER_LEFT_1_END_STABILITY=6.9000
PRIMER_RIGHT_1_END_STABILITY=6.4000
PRIMER_PAIR_1_COMPL_ANY=7.00
PRIMER_PAIR_1_COMPL_END=2.00
PRIMER_PRODUCT_SIZE_1=172
PRIMER_PAIR_PENALTY_2=1.2615
PRIMER_LEFT_2_PENALTY=0.243965
PRIMER_RIGHT_2_PENALTY=1.017510
PRIMER_INTERNAL_OLIGO_2_PENALTY=0.038050
PRIMER_LEFT_2_SEQUENCE=ACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_2_SEQUENCE=AATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_2_SEQUENCE=AAAAAAAAAAAAAAAAAAAAAAAAA
PRIMER_LEFT_2=3,24
PRIMER_RIGHT_2=172,24
PRIMER_INTERNAL_OLIGO_2=61,25
PRIMER_LEFT_2_TM=60.244
PRIMER_RIGHT_2_TM=58.982
PRIMER_INTERNAL_OLIGO_2_TM=55.838
PRIMER_LEFT_2_GC_PERCENT=33.333
PRIMER_RIGHT_2_GC_PERCENT=33.333
PRIMER_INTERNAL_OLIGO_2_GC_PERCENT=0.000
PRIMER_LEFT_2_SELF_ANY=12.00
PRIMER_RIGHT_2_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_2_SELF_ANY=0.00
PRIMER_LEFT_2_SELF_END=10.00
PRIMER_RIGHT_2_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_2_SELF_END=0.00
PRIMER_LEFT_2_END_STABILITY=6.9000
PRIMER_RIGHT_2_END_STABILITY=6.4000
PRIMER_PAIR_2_COMPL_ANY=6.00
PRIMER_PAIR_2_COMPL_END=1.00
PRIMER_PRODUCT_SIZE_2=170
PRIMER_PAIR_PENALTY_3=1.2615
PRIMER_LEFT_3_PENALTY=0.243965
PRIMER_RIGHT_3_PENALTY=1.017510
PRIMER_INTERNAL_OLIGO_3_PENALTY=0.038050
PRIMER_LEFT_3_SEQUENCE=CACGTCATCTCATCAAATTTGATT
PRIMER_RIGHT_3_SEQUENCE=AATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_3_SEQUENCE=AAAAAAAAAAAAAAAAAAAAAAAAA
PRIMER_LEFT_3=2,24
PRIMER_RIGHT_3=172,24
PRIMER_INTERNAL_OLIGO_3=61,25
PRIMER_LEFT_3_TM=60.244
PRIMER_RIGHT_3_TM=58.982
PRIMER_INTERNAL_OLIGO_3_TM=55.838
PRIMER_LEFT_3_GC_PERCENT=33.333
PRIMER_RIGHT_3_GC_PERCENT=33.333
PRIMER_INTERNAL_OLIGO_3_GC_PERCENT=0.000
PRIMER_LEFT_3_SELF_ANY=12.00
PRIMER_RIGHT_3_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_3_SELF_ANY=0.00
PRIMER_LEFT_3_SELF_END=10.00
PRIMER_RIGHT_3_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_3_SELF_END=0.00
PRIMER_LEFT_3_END_STABILITY=6.9000
PRIMER_RIGHT_3_END_STABILITY=6.4000
PRIMER_PAIR_3_COMPL_ANY=7.00
PRIMER_PAIR_3_COMPL_END=2.00
PRIMER_PRODUCT_SIZE_3=171
PRIMER_PAIR_PENALTY_4=29.3957
PRIMER_LEFT_4_PENALTY=29.308664
PRIMER_RIGHT_4_PENALTY=0.087031
PRIMER_INTERNAL_OLIGO_4_PENALTY=0.038050
PRIMER_LEFT_4_SEQUENCE=GGGGGGGGGGGGGGGGGGGGGGGG
PRIMER_RIGHT_4_SEQUENCE=GAATTGTTTGGAAATGACAGTGAG
PRIMER_INTERNAL_OLIGO_4_SEQUENCE=AAAAAAAAAAAAAAAAAAAAAAAAA
PRIMER_LEFT_4=37,24
PRIMER_RIGHT_4=173,24
PRIMER_INTERNAL_OLIGO_4=61,25
PRIMER_LEFT_4_TM=89.309
PRIMER_RIGHT_4_TM=59.913
PRIMER_INTERNAL_OLIGO_4_TM=55.838
PRIMER_LEFT_4_GC_PERCENT=100.000
PRIMER_RIGHT_4_GC_PERCENT=37.500
PRIMER_INTERNAL_OLIGO_4_GC_PERCENT=0.000
PRIMER_LEFT_4_SELF_ANY=0.00
PRIMER_RIGHT_4_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_4_SELF_ANY=0.00
PRIMER_LEFT_4_SELF_END=0.00
PRIMER_RIGHT_4_SELF_END=1.00
PRIMER_INTERNAL_OLIGO_4_SELF_END=0.00
PRIMER_LEFT_4_END_STABILITY=12.4000
PRIMER_RIGHT_4_END_STABILITY=6.4000
PRIMER_PAIR_4_COMPL_ANY=1.00
PRIMER_PAIR_4_COMPL_END=0.00
PRIMER_PRODUCT_SIZE_4=137
=
MARKER_NAME=boundary_test_22
PRIMER_EXPLAIN_FLAG=1
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
PRIMER_PRODUCT_SIZE_RANGE=80-80
PRIMER_MIN_TM=0.0
PRIMER_MAX_TM=100.0
PRIMER_MIN_SIZE=25
PRIMER_MAX_SIZE=25
PRIMER_INTERNAL_OLIGO_MIN_SIZE=25
PRIMER_INTERNAL_OLIGO_MAX_SIZE=25
PRIMER_INTERNAL_OLIGO_OPT_SIZE=25
PRIMER_OPT_SIZE=25
PRIMER_MAX_GC=100
PRIMER_MIN_GC=0
PRIMER_SELF_END=100
PRIMER_SELF_ANY=100
PRIMER_INTERNAL_OLIGO_SELF_ANY=100
PRIMER_INTERNAL_OLIGO_SELF_END=100 
PRIMER_INTERNAL_OLIGO_MAX_TM=100
PRIMER_INTERNAL_OLIGO_MIN_TM=30
PRIMER_INTERNAL_OLIGO_MAX_GC=100
PRIMER_INTERNAL_OLIGO_MIN_GC=0
PRIMER_PICK_INTERNAL_OLIGO=1
COMMENT=should succeed
PRIMER_LEFT_EXPLAIN=considered 5, too many Ns 4, ok 1
PRIMER_RIGHT_EXPLAIN=considered 5, too many Ns 4, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 60, too many Ns 57, ok 3
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=15.695378
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=ACGTACGTACGTTGCAGTCGACGCA
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=30,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=71.495
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=56.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=8.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_23
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
INCLUDED_REGION=2,80
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=1,1
COMMENT=should succeed with warning (internal oligo excluded region outside of included region)
PRIMER_WARNING=PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION outside of INCLUDED_REGION
PRIMER_LEFT_EXPLAIN=considered 1, ok 1
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 56, too many Ns 53, ok 3
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=15.695378
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=ACGTACGTACGTTGCAGTCGACGCA
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=30,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=71.495
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=56.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=8.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_24
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
INCLUDED_REGION=2,80
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=2,1
COMMENT=should succeed
PRIMER_LEFT_EXPLAIN=considered 1, ok 1
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 56, too many Ns 53, overlap excluded region 1, ok 2
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=15.695378
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=ACGTACGTACGTTGCAGTCGACGCA
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=30,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=71.495
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=56.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=8.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_25
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
INCLUDED_REGION=2,80
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=0,3
COMMENT=should succeed with warning (internal oligo excluded region outside of included region)
PRIMER_WARNING=PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION outside of INCLUDED_REGION
PRIMER_LEFT_EXPLAIN=considered 1, ok 1
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 56, too many Ns 53, overlap excluded region 1, ok 2
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=15.695378
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=ACGTACGTACGTTGCAGTCGACGCA
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=30,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=71.495
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=56.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=8.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_26
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
INCLUDED_REGION=2,80
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=29,1
COMMENT=should succeed
PRIMER_LEFT_EXPLAIN=considered 1, ok 1
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 56, too many Ns 53, ok 3
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=15.695378
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=ACGTACGTACGTTGCAGTCGACGCA
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=30,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=71.495
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=56.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=8.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_27
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
INCLUDED_REGION=2,80
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=30,1
COMMENT=should fail w/ no legal internal oligo
PRIMER_LEFT_EXPLAIN=considered 1, ok 1
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 56, too many Ns 53, overlap excluded region 1, ok 2
PRIMER_PAIR_EXPLAIN=considered 1, no internal oligo 1, ok 0
=
MARKER_NAME=boundary_test_28
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
INCLUDED_REGION=2,80
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=54,1
COMMENT=should fail w/ no legal internal oligo
PRIMER_LEFT_EXPLAIN=considered 1, ok 1
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 56, too many Ns 53, overlap excluded region 1, ok 2
PRIMER_PAIR_EXPLAIN=considered 1, no internal oligo 1, ok 0
=
MARKER_NAME=boundary_test_29
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
INCLUDED_REGION=2,80
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=55,1
COMMENT=should succeed
PRIMER_LEFT_EXPLAIN=considered 1, ok 1
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 56, too many Ns 53, ok 3
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=15.695378
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=ACGTACGTACGTTGCAGTCGACGCA
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=30,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=71.495
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=56.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=8.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_30
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
INCLUDED_REGION=2,80
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=81,1
COMMENT=should succeed
PRIMER_LEFT_EXPLAIN=considered 1, ok 1
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 56, too many Ns 53, overlap excluded region 1, ok 2
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=15.695378
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=ACGTACGTACGTTGCAGTCGACGCA
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=30,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=71.495
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=56.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=8.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_31
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
INCLUDED_REGION=2,80
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=82,1
COMMENT=should generate warning (external oligo excluded region outside of included region)
PRIMER_WARNING=PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION outside of INCLUDED_REGION
PRIMER_LEFT_EXPLAIN=considered 1, ok 1
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 56, too many Ns 53, ok 3
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=15.695378
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=ACGTACGTACGTTGCAGTCGACGCA
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=30,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=71.495
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=56.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=8.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_32
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
INCLUDED_REGION=2,80
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=81,2
COMMENT=should generate warning (external oligo excluded region outside of included region)
PRIMER_WARNING=PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION outside of INCLUDED_REGION
PRIMER_LEFT_EXPLAIN=considered 1, ok 1
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 56, too many Ns 53, overlap excluded region 1, ok 2
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=15.695378
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=ACGTACGTACGTTGCAGTCGACGCA
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=30,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=71.495
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=56.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=8.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_33
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
INCLUDED_REGION=2,80
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1
COMMENT=should succeed (testing number of internal oligo excluded regions)
PRIMER_LEFT_EXPLAIN=considered 1, ok 1
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_INTERNAL_OLIGO_EXPLAIN=considered 56, too many Ns 53, overlap excluded region 1, ok 2
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=3.3813
PRIMER_LEFT_PENALTY=2.731673
PRIMER_RIGHT_PENALTY=0.649600
PRIMER_INTERNAL_OLIGO_PENALTY=15.695378
PRIMER_LEFT_SEQUENCE=CACGTCATCTCATCAAATTTGATTG
PRIMER_RIGHT_SEQUENCE=GAATTGTTTGGAAATGACAGTGAGT
PRIMER_INTERNAL_OLIGO_SEQUENCE=ACGTACGTACGTTGCAGTCGACGCA
PRIMER_LEFT=2,25
PRIMER_RIGHT=81,25
PRIMER_INTERNAL_OLIGO=30,25
PRIMER_LEFT_TM=62.732
PRIMER_RIGHT_TM=60.650
PRIMER_INTERNAL_OLIGO_TM=71.495
PRIMER_LEFT_GC_PERCENT=36.000
PRIMER_RIGHT_GC_PERCENT=36.000
PRIMER_INTERNAL_OLIGO_GC_PERCENT=56.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_INTERNAL_OLIGO_SELF_ANY=12.00
PRIMER_LEFT_SELF_END=10.00
PRIMER_RIGHT_SELF_END=3.00
PRIMER_INTERNAL_OLIGO_SELF_END=8.00
PRIMER_LEFT_END_STABILITY=6.9000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_PAIR_COMPL_ANY=7.00
PRIMER_PAIR_COMPL_END=1.00
PRIMER_PRODUCT_SIZE=80
=
MARKER_NAME=boundary_test_34
SEQUENCE=NNCACGTCATCTCATCAAATTTGATTGNNNACGTACGTACGTTGCAGTCGACGCANNACTCACTGTCATTTCCAAACAATTCNN
INCLUDED_REGION=2,80
PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION=81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1 81,1
COMMENT=should fail (testing number of internal oligo excluded regions)
PRIMER_ERROR=Too many elements for tag PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION
=
