PRIMER_SEQUENCE_ID=template_mis_1
PRIMER_NUM_RETURN=1
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_PAIR_MAX_TEMPLATE_MISPRIMING=100
SEQUENCE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
PRIMER_PAIR_PENALTY=0.9341
PRIMER_LEFT_PENALTY=0.323851
PRIMER_RIGHT_PENALTY=0.610234
PRIMER_LEFT_SEQUENCE=Gatgcgtacgatccatgcta
PRIMER_RIGHT_SEQUENCE=tagcggcctagtgtgtgtgt
PRIMER_LEFT=20,20
PRIMER_RIGHT=131,20
PRIMER_LEFT_TM=59.676
PRIMER_RIGHT_TM=59.390
PRIMER_LEFT_GC_PERCENT=50.000
PRIMER_RIGHT_GC_PERCENT=55.000
PRIMER_LEFT_SELF_ANY=8.00
PRIMER_RIGHT_SELF_ANY=4.00
PRIMER_LEFT_SELF_END=2.00
PRIMER_RIGHT_SELF_END=0.00
PRIMER_LEFT_END_STABILITY=7.5000
PRIMER_RIGHT_END_STABILITY=6.4000
PRIMER_LEFT_TEMPLATE_MISPRIMING=6.0000
PRIMER_RIGHT_TEMPLATE_MISPRIMING=6.0000
PRIMER_PAIR_COMPL_ANY=4.00
PRIMER_PAIR_COMPL_END=2.00
PRIMER_PRODUCT_SIZE=112
PRIMER_PAIR_TEMPLATE_MISPRIMING=12.00
=
PRIMER_SEQUENCE_ID=template_mis_2
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_LEFT_INPUT=TCCTACAGCTGTGGGAAAATGat
PRIMER_PAIR_MAX_TEMPLATE_MISPRIMING=100
PRIMER_PAIR_WT_TEMPLATE_MISPRIMING=1
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
SEQUENCE=TCCTACAGCTGTGGGAAAATGatgcgtacgatTCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTGatatcatctagcGCCGCCATGTACACACTG
PRIMER_WARNING=Left primer is unacceptable: High self complementarity
PRIMER_RIGHT_EXPLAIN=considered 1240, low tm 698, high tm 374, high any compl 13, high end compl 34, ok 121
PRIMER_PAIR_EXPLAIN=considered 242, ok 242
PRIMER_PAIR_PENALTY=2600.1166
PRIMER_LEFT_PENALTY=0.102704
PRIMER_RIGHT_PENALTY=0.013916
PRIMER_LEFT_SEQUENCE=TCCTACAGCTGTGGGAAAATGat
PRIMER_RIGHT_SEQUENCE=gatatCAGTGTGTACATGGCGG
PRIMER_LEFT=32,23
PRIMER_RIGHT=197,22
PRIMER_LEFT_TM=62.054
PRIMER_RIGHT_TM=60.278
PRIMER_LEFT_GC_PERCENT=43.478
PRIMER_RIGHT_GC_PERCENT=50.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=6.00
PRIMER_LEFT_SELF_END=3.00
PRIMER_RIGHT_SELF_END=1.00
PRIMER_LEFT_END_STABILITY=6.5000
PRIMER_RIGHT_END_STABILITY=12.9000
PRIMER_LEFT_TEMPLATE_MISPRIMING=23.0000
PRIMER_RIGHT_TEMPLATE_MISPRIMING=17.0000
PRIMER_PAIR_COMPL_ANY=4.00
PRIMER_PAIR_COMPL_END=2.00
PRIMER_PRODUCT_SIZE=166
PRIMER_PAIR_TEMPLATE_MISPRIMING=26.00
=
PRIMER_SEQUENCE_ID=template_mis_3
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_LEFT_INPUT=TCCTACAGCTGTGGGAAAATGat
PRIMER_RIGHT_INPUT=GGGGGGGGGGGGGGGGGG
PRIMER_PAIR_MAX_TEMPLATE_MISPRIMING=100
PRIMER_PAIR_WT_TEMPLATE_MISPRIMING=1
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
SEQUENCE=TCCTACAGCTGTGGGAAAATGatgcgtacgatTCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTGatatcatctagcGCCGCCATGTACACACTGCCCCCCCCCCCCCCCCCC
PRIMER_WARNING=Left primer is unacceptable: High self complementarity; Right primer is unacceptable: Unacceptable GC content/Tm too high/Long poly-X
PRIMER_PAIR_EXPLAIN=considered 2, ok 2
PRIMER_PAIR_PENALTY=2601.0969
PRIMER_LEFT_PENALTY=0.102704
PRIMER_RIGHT_PENALTY=0.994170
PRIMER_LEFT_SEQUENCE=TCCTACAGCTGTGGGAAAATGat
PRIMER_RIGHT_SEQUENCE=GGGGGGGGGGGGGGGGGG
PRIMER_LEFT=32,23
PRIMER_RIGHT=240,18
PRIMER_LEFT_TM=62.054
PRIMER_RIGHT_TM=79.883
PRIMER_LEFT_GC_PERCENT=43.478
PRIMER_RIGHT_GC_PERCENT=100.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=0.00
PRIMER_LEFT_SELF_END=3.00
PRIMER_RIGHT_SELF_END=0.00
PRIMER_LEFT_END_STABILITY=6.5000
PRIMER_RIGHT_END_STABILITY=12.4000
PRIMER_LEFT_TEMPLATE_MISPRIMING=23.0000
PRIMER_RIGHT_TEMPLATE_MISPRIMING=3.0000
PRIMER_PAIR_COMPL_ANY=2.00
PRIMER_PAIR_COMPL_END=0.00
PRIMER_PRODUCT_SIZE=209
PRIMER_PAIR_TEMPLATE_MISPRIMING=26.00
=
PRIMER_SEQUENCE_ID=template_mis_4
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_LEFT_INPUT=TCCTACAGCTGTGGGAAAATGat
PRIMER_RIGHT_INPUT=GGGGGGGGGGGGGGGGGG
PRIMER_PAIR_MAX_TEMPLATE_MISPRIMING=100
PRIMER_PAIR_WT_TEMPLATE_MISPRIMING=1
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_LIBERAL_BASE=1
SEQUENCE=TCCTACAGCTGTGGGAAAATGatTCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTGatatcatctagcGCCGCCATGTACACACTGCCCCCCCCCCCCCCCCCCACSCCCCCCCCCCCCCCCCA
PRIMER_WARNING=Unrecognized base in input sequence; Left primer is unacceptable: High self complementarity; Right primer is unacceptable: Unacceptable GC content/Tm too high/Long poly-X
PRIMER_PAIR_EXPLAIN=considered 2, ok 2
PRIMER_PAIR_PENALTY=2701.0969
PRIMER_LEFT_PENALTY=0.102704
PRIMER_RIGHT_PENALTY=0.994170
PRIMER_LEFT_SEQUENCE=TCCTACAGCTGTGGGAAAATGat
PRIMER_RIGHT_SEQUENCE=GGGGGGGGGGGGGGGGGG
PRIMER_LEFT=23,23
PRIMER_RIGHT=231,18
PRIMER_LEFT_TM=62.054
PRIMER_RIGHT_TM=79.883
PRIMER_LEFT_GC_PERCENT=43.478
PRIMER_RIGHT_GC_PERCENT=100.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=0.00
PRIMER_LEFT_SELF_END=3.00
PRIMER_RIGHT_SELF_END=0.00
PRIMER_LEFT_END_STABILITY=6.5000
PRIMER_RIGHT_END_STABILITY=12.4000
PRIMER_LEFT_TEMPLATE_MISPRIMING=23.0000
PRIMER_RIGHT_TEMPLATE_MISPRIMING=16.0000
PRIMER_PAIR_COMPL_ANY=2.00
PRIMER_PAIR_COMPL_END=0.00
PRIMER_PRODUCT_SIZE=209
PRIMER_PAIR_TEMPLATE_MISPRIMING=27.00
=
PRIMER_SEQUENCE_ID=template_mis_5
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_LEFT_INPUT=TCCTACAGCTGTGGGAAAATGat
PRIMER_RIGHT_INPUT=GGGGGGGGGGGGGGGGGG
PRIMER_PAIR_MAX_TEMPLATE_MISPRIMING=100
PRIMER_PAIR_WT_TEMPLATE_MISPRIMING=1
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_LIBERAL_BASE=1
SEQUENCE=TCCTACAGCTGTGGGAAAATGatTCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTGatatcatctagcGCCGCCATGTACACACTGCCCCCCCCCCCCCCCCCCACTCCCCCCCCCCCCCCCCA
PRIMER_WARNING=Left primer is unacceptable: High self complementarity; Right primer is unacceptable: Unacceptable GC content/Tm too high/Long poly-X
PRIMER_PAIR_EXPLAIN=considered 2, ok 2
PRIMER_PAIR_PENALTY=2701.0969
PRIMER_LEFT_PENALTY=0.102704
PRIMER_RIGHT_PENALTY=0.994170
PRIMER_LEFT_SEQUENCE=TCCTACAGCTGTGGGAAAATGat
PRIMER_RIGHT_SEQUENCE=GGGGGGGGGGGGGGGGGG
PRIMER_LEFT=23,23
PRIMER_RIGHT=231,18
PRIMER_LEFT_TM=62.054
PRIMER_RIGHT_TM=79.883
PRIMER_LEFT_GC_PERCENT=43.478
PRIMER_RIGHT_GC_PERCENT=100.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=0.00
PRIMER_LEFT_SELF_END=3.00
PRIMER_RIGHT_SELF_END=0.00
PRIMER_LEFT_END_STABILITY=6.5000
PRIMER_RIGHT_END_STABILITY=12.4000
PRIMER_LEFT_TEMPLATE_MISPRIMING=23.0000
PRIMER_RIGHT_TEMPLATE_MISPRIMING=16.0000
PRIMER_PAIR_COMPL_ANY=2.00
PRIMER_PAIR_COMPL_END=0.00
PRIMER_PRODUCT_SIZE=209
PRIMER_PAIR_TEMPLATE_MISPRIMING=27.00
=
PRIMER_SEQUENCE_ID=template_mis_6
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_LEFT_INPUT=TCCTACAGCTGTGGGAAAATGat
PRIMER_RIGHT_INPUT=GGGGGGGGGGGGGGGGGG
PRIMER_PAIR_MAX_TEMPLATE_MISPRIMING=100
PRIMER_PAIR_WT_TEMPLATE_MISPRIMING=1
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_LIBERAL_BASE=1
PRIMER_PRODUCT_SIZE_RANGE=36-300
SEQUENCE=TCCTACAGCTGTGGGAAAATGatNgcgtacNCCCCCCCCCACCCCCCCCACCCCCCCCCCCCCCCCCC
PRIMER_WARNING=Left primer is unacceptable: High self complementarity; Right primer is unacceptable: Unacceptable GC content/Tm too high/Long poly-X
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=1901.0969
PRIMER_LEFT_PENALTY=0.102704
PRIMER_RIGHT_PENALTY=0.994170
PRIMER_LEFT_SEQUENCE=TCCTACAGCTGTGGGAAAATGat
PRIMER_RIGHT_SEQUENCE=GGGGGGGGGGGGGGGGGG
PRIMER_LEFT=0,23
PRIMER_RIGHT=67,18
PRIMER_LEFT_TM=62.054
PRIMER_RIGHT_TM=79.883
PRIMER_LEFT_GC_PERCENT=43.478
PRIMER_RIGHT_GC_PERCENT=100.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=0.00
PRIMER_LEFT_SELF_END=3.00
PRIMER_RIGHT_SELF_END=0.00
PRIMER_LEFT_END_STABILITY=6.5000
PRIMER_RIGHT_END_STABILITY=12.4000
PRIMER_LEFT_TEMPLATE_MISPRIMING=3.0000
PRIMER_RIGHT_TEMPLATE_MISPRIMING=16.0000
PRIMER_PAIR_COMPL_ANY=2.00
PRIMER_PAIR_COMPL_END=0.00
PRIMER_PRODUCT_SIZE=68
PRIMER_PAIR_TEMPLATE_MISPRIMING=19.00
=
PRIMER_SEQUENCE_ID=template_mis_7
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_LEFT_INPUT=TCCTACAGCTGTGGGAAAATGat
PRIMER_RIGHT_INPUT=GGGGGGGGGGGGGGGGGG
PRIMER_PAIR_MAX_TEMPLATE_MISPRIMING=100
PRIMER_PAIR_WT_TEMPLATE_MISPRIMING=1
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_LIBERAL_BASE=1
PRIMER_PRODUCT_SIZE_RANGE=36-300
SEQUENCE=TCCTACAGCTGTGGGAAAATGatNgcgtacNCCCCCCCCCAAAAAAAAAACCCCCCCCCCCCCCCCCC
PRIMER_WARNING=Left primer is unacceptable: High self complementarity; Right primer is unacceptable: Unacceptable GC content/Tm too high/Long poly-X
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_PENALTY=1276.0969
PRIMER_LEFT_PENALTY=0.102704
PRIMER_RIGHT_PENALTY=0.994170
PRIMER_LEFT_SEQUENCE=TCCTACAGCTGTGGGAAAATGat
PRIMER_RIGHT_SEQUENCE=GGGGGGGGGGGGGGGGGG
PRIMER_LEFT=0,23
PRIMER_RIGHT=67,18
PRIMER_LEFT_TM=62.054
PRIMER_RIGHT_TM=79.883
PRIMER_LEFT_GC_PERCENT=43.478
PRIMER_RIGHT_GC_PERCENT=100.000
PRIMER_LEFT_SELF_ANY=12.00
PRIMER_RIGHT_SELF_ANY=0.00
PRIMER_LEFT_SELF_END=3.00
PRIMER_RIGHT_SELF_END=0.00
PRIMER_LEFT_END_STABILITY=6.5000
PRIMER_RIGHT_END_STABILITY=12.4000
PRIMER_LEFT_TEMPLATE_MISPRIMING=3.0000
PRIMER_RIGHT_TEMPLATE_MISPRIMING=9.7500
PRIMER_PAIR_COMPL_ANY=2.00
PRIMER_PAIR_COMPL_END=0.00
PRIMER_PRODUCT_SIZE=68
PRIMER_PAIR_TEMPLATE_MISPRIMING=12.75
=
PRIMER_SEQUENCE_ID=template_mis_8
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_TEMPLATE_MISPRIMING=11
PRIMER_MAX_POLY_X=40
PRIMER_PAIR_MAX_TEMPLATE_MISPRIMING=100
PRIMER_PAIR_WT_TEMPLATE_MISPRIMING=1
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_LIBERAL_BASE=1
PRIMER_PRODUCT_SIZE_RANGE=36-300
PRIMER_MAX_GC=100
PRIMER_MAX_TM=100
PRIMER_RIGHT_INPUT=GGGGGGGGGGGGGGGGGG
SEQUENCE=TCCTACAGCTGTGGGAAAATGatNgcgtacNCCCCCCCCCAAAAAAAAAACCCCCCCCCCCCCCCCCC
COMMENT=Should succeed
PRIMER_LEFT_EXPLAIN=considered 75, too many Ns 34, low tm 14, high any compl 7, ok 20
PRIMER_PAIR_EXPLAIN=considered 20, high any compl 11, high end compl 5, ok 4
PRIMER_PAIR_PENALTY=1076.0792
PRIMER_LEFT_PENALTY=0.085063
PRIMER_RIGHT_PENALTY=0.994170
PRIMER_LEFT_SEQUENCE=CCCCCCCCAAAAAAAAAA
PRIMER_RIGHT_SEQUENCE=GGGGGGGGGGGGGGGGGG
PRIMER_LEFT=32,18
PRIMER_RIGHT=67,18
PRIMER_LEFT_TM=61.701
PRIMER_RIGHT_TM=79.883
PRIMER_LEFT_GC_PERCENT=44.444
PRIMER_RIGHT_GC_PERCENT=100.000
PRIMER_LEFT_SELF_ANY=0.00
PRIMER_RIGHT_SELF_ANY=0.00
PRIMER_LEFT_SELF_END=0.00
PRIMER_RIGHT_SELF_END=0.00
PRIMER_LEFT_END_STABILITY=7.6000
PRIMER_RIGHT_END_STABILITY=12.4000
PRIMER_LEFT_TEMPLATE_MISPRIMING=4.0000
PRIMER_RIGHT_TEMPLATE_MISPRIMING=9.7500
PRIMER_PAIR_COMPL_ANY=8.00
PRIMER_PAIR_COMPL_END=0.00
PRIMER_PRODUCT_SIZE=36
PRIMER_PAIR_TEMPLATE_MISPRIMING=10.75
=
PRIMER_SEQUENCE_ID=template_mis_9
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_TEMPLATE_MISPRIMING=9
PRIMER_MAX_POLY_X=40
PRIMER_SELF_END=100
PRIMER_PAIR_MAX_TEMPLATE_MISPRIMING=100
PRIMER_PAIR_WT_TEMPLATE_MISPRIMING=1
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_LIBERAL_BASE=1
PRIMER_PRODUCT_SIZE_RANGE=36-300
PRIMER_MAX_GC=100
SEQUENCE=TCCTACAGCTGTGGGAAAATGatNgcgtacNCCCCCCCCCAAAAAAAAAACCCCCCCCCCCCCCCCCC
COMMENT=Should succeed
PRIMER_LEFT_EXPLAIN=considered 75, too many Ns 34, low tm 14, high any compl 7, ok 20
PRIMER_RIGHT_EXPLAIN=considered 177, too many Ns 22,high template mispriming score 5, ok 150
PRIMER_PAIR_EXPLAIN=considered 3004, unacceptable product size 2784, high any compl 199, ok 21
PRIMER_PAIR_PENALTY=776.1163
PRIMER_LEFT_PENALTY=0.085063
PRIMER_RIGHT_PENALTY=1.031264
PRIMER_LEFT_SEQUENCE=CCCCCCCCAAAAAAAAAA
PRIMER_RIGHT_SEQUENCE=GGGGGGGGGGGGGGGGGGTTTTT
PRIMER_LEFT=32,18
PRIMER_RIGHT=67,23
PRIMER_LEFT_TM=61.701
PRIMER_RIGHT_TM=80.625
PRIMER_LEFT_GC_PERCENT=44.444
PRIMER_RIGHT_GC_PERCENT=78.261
PRIMER_LEFT_SELF_ANY=0.00
PRIMER_RIGHT_SELF_ANY=0.00
PRIMER_LEFT_SELF_END=0.00
PRIMER_RIGHT_SELF_END=0.00
PRIMER_LEFT_END_STABILITY=7.6000
PRIMER_RIGHT_END_STABILITY=7.6000
PRIMER_LEFT_TEMPLATE_MISPRIMING=4.0000
PRIMER_RIGHT_TEMPLATE_MISPRIMING=6.7500
PRIMER_PAIR_COMPL_ANY=8.00
PRIMER_PAIR_COMPL_END=5.00
PRIMER_PRODUCT_SIZE=36
PRIMER_PAIR_TEMPLATE_MISPRIMING=7.75
=
PRIMER_SEQUENCE_ID=template_mis_10
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_TEMPLATE_MISPRIMING=6.7
PRIMER_MAX_POLY_X=40
PRIMER_SELF_END=100
PRIMER_PAIR_MAX_TEMPLATE_MISPRIMING=100
PRIMER_PAIR_WT_TEMPLATE_MISPRIMING=1
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_LIBERAL_BASE=1
PRIMER_PRODUCT_SIZE_RANGE=36-300
PRIMER_MAX_GC=100
SEQUENCE=TCCTACAGCTGTGGGAAAATGatNgcgtacNCCCCCCCCCAAAAAAAAAACCCCCCCCCCCCCCCCCC
COMMENT=Should fail
PRIMER_LEFT_EXPLAIN=considered 75, too many Ns 34, low tm 14, high any compl 7,high template mispriming score 2, ok 18
PRIMER_RIGHT_EXPLAIN=considered 177, too many Ns 22,high template mispriming score 19, ok 136
PRIMER_PAIR_EXPLAIN=considered 2718, unacceptable product size 2710, high any compl 8, ok 0
=
PRIMER_SEQUENCE_ID=template_mis_11
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_POLY_X=40
PRIMER_SELF_END=100
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_MAX_GC=100
PRIMER_MAX_TM=100
PRIMER_TASK=pick_left_only
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_LEFT_INPUT=cTTGATGGAGAATTGCGCAGAGGGGc
SEQUENCE=aTTGATGGAGAATTGCGCAGAGGGGcTTGATGGAGAATTGCGCAGAGGGGc
COMMENT=Test off-by-one error in checking for mispriming on the same strand as the oligo
PRIMER_LEFT_PENALTY=0.7321
PRIMER_LEFT_SEQUENCE=cTTGATGGAGAATTGCGCAGAGGGGc
PRIMER_LEFT=25,26
PRIMER_LEFT_TM=74.643
PRIMER_LEFT_GC_PERCENT=57.692
PRIMER_LEFT_SELF_ANY=6.00
PRIMER_LEFT_SELF_END=2.00
PRIMER_LEFT_END_STABILITY=12.4000
PRIMER_LEFT_TEMPLATE_MISPRIMING=20.00
=
PRIMER_SEQUENCE_ID=template_mis_12
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_POLY_X=40
PRIMER_SELF_END=100
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_MAX_GC=100
PRIMER_MAX_TM=100
PRIMER_TASK=pick_left_only
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_LEFT_INPUT=cTTGATGGAGAATTGCGCAGAGGGGc
INCLUDED_REGION=25,26
SEQUENCE=aTTGATGGAGAATTGCGCAGAGGGGcTTGATGGAGAATTGCGCAGAGGGGc
COMMENT=Test off-by-one error in checking for mispriming on the same strand as the oligo plus former error in ignoring included_region offset
PRIMER_LEFT_PENALTY=0.7321
PRIMER_LEFT_SEQUENCE=cTTGATGGAGAATTGCGCAGAGGGGc
PRIMER_LEFT=25,26
PRIMER_LEFT_TM=74.643
PRIMER_LEFT_GC_PERCENT=57.692
PRIMER_LEFT_SELF_ANY=6.00
PRIMER_LEFT_SELF_END=2.00
PRIMER_LEFT_END_STABILITY=12.4000
PRIMER_LEFT_TEMPLATE_MISPRIMING=20.00
=
PRIMER_SEQUENCE_ID=template_mis_13
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_POLY_X=40
PRIMER_SELF_END=100
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_MAX_GC=100
PRIMER_MAX_TM=100
PRIMER_TASK=pick_left_only
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_LEFT_INPUT=cTTGATGGAGAATTGCGCAGAGGGGc
SEQUENCE=cTTGATGGAGAATTGCGCAGAGGGGcTTGATGGAGAATTGCGCAGAGGGc
COMMENT=Test off-by-one error in checking for mispriming on the same strand as the oligo
PRIMER_LEFT_PENALTY=0.7321
PRIMER_LEFT_SEQUENCE=cTTGATGGAGAATTGCGCAGAGGGGc
PRIMER_LEFT=0,26
PRIMER_LEFT_TM=74.643
PRIMER_LEFT_GC_PERCENT=57.692
PRIMER_LEFT_SELF_ANY=6.00
PRIMER_LEFT_SELF_END=2.00
PRIMER_LEFT_END_STABILITY=12.4000
PRIMER_LEFT_TEMPLATE_MISPRIMING=22.00
=
PRIMER_SEQUENCE_ID=template_mis_14
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_POLY_X=40
PRIMER_SELF_END=100
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_MAX_GC=100
PRIMER_MAX_TM=100
PRIMER_TASK=pick_right_only
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_RIGHT_INPUT=cTTGATGGAGAATTGCGCAGAGGGGc
SEQUENCE=GCCCCTCTGCGCAATTCTCCATCAAGCCCCTCTGCGCAATTCTCCATCAAT
COMMENT=Test off-by-one error in checking for mispriming on the same strand as the oligo; SEQUENCE is complements of template_mis_11
PRIMER_RIGHT_PENALTY=0.7321
PRIMER_RIGHT_SEQUENCE=CTTGATGGAGAATTGCGCAGAGGGGC
PRIMER_RIGHT=25,26
PRIMER_RIGHT_TM=74.643
PRIMER_RIGHT_GC_PERCENT=57.692
PRIMER_RIGHT_SELF_ANY=6.00
PRIMER_RIGHT_SELF_END=2.00
PRIMER_RIGHT_END_STABILITY=12.4000
PRIMER_RIGHT_TEMPLATE_MISPRIMING=20.00
=
PRIMER_SEQUENCE_ID=template_mis_15
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_POLY_X=40
PRIMER_SELF_END=100
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_MAX_GC=100
PRIMER_MAX_TM=100
PRIMER_TASK=pick_right_only
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_RIGHT_INPUT=cTTGATGGAGAATTGCGCAGAGGGGc
SEQUENCE=GCCCTCTGCGCAATTCTCCATCAAGCCCCTCTGCGCAATTCTCCATCAAG
COMMENT=Test off-by-one error in checking for mispriming on the same strand as the oligo; SEQUENCE is complements of template_mis_13
PRIMER_RIGHT_PENALTY=0.7321
PRIMER_RIGHT_SEQUENCE=CTTGATGGAGAATTGCGCAGAGGGGC
PRIMER_RIGHT=49,26
PRIMER_RIGHT_TM=74.643
PRIMER_RIGHT_GC_PERCENT=57.692
PRIMER_RIGHT_SELF_ANY=6.00
PRIMER_RIGHT_SELF_END=2.00
PRIMER_RIGHT_END_STABILITY=12.4000
PRIMER_RIGHT_TEMPLATE_MISPRIMING=22.00
=
PRIMER_SEQUENCE_ID=template_mis_16
PRIMER_NUM_RETURN=1
PRIMER_EXPLAIN_FLAG=1
PRIMER_PICK_ANYWAY=1
PRIMER_MAX_POLY_X=40
PRIMER_SELF_END=100
PRIMER_WT_SIZE_GT=0
PRIMER_WT_SIZE_LT=0
PRIMER_WT_TM_GT=.05
PRIMER_WT_TM_LT=.05
PRIMER_MAX_GC=100
PRIMER_MAX_TM=100
PRIMER_TASK=pick_right_only
PRIMER_MAX_TEMPLATE_MISPRIMING=100
PRIMER_RIGHT_INPUT=cTTGATGGAGAATTGCGCAGAGGGGc
INCLUDED_REGION=24,26
SEQUENCE=GCCCTCTGCGCAATTCTCCATCAAGCCCCTCTGCGCAATTCTCCATCAAG
COMMENT=Test off-by-one error in checking for mispriming on the same strand as the oligo; SEQUENCE is complements of template_mis_13
PRIMER_RIGHT_PENALTY=0.7321
PRIMER_RIGHT_SEQUENCE=CTTGATGGAGAATTGCGCAGAGGGGC
PRIMER_RIGHT=49,26
PRIMER_RIGHT_TM=74.643
PRIMER_RIGHT_GC_PERCENT=57.692
PRIMER_RIGHT_SELF_ANY=6.00
PRIMER_RIGHT_SELF_END=2.00
PRIMER_RIGHT_END_STABILITY=12.4000
PRIMER_RIGHT_TEMPLATE_MISPRIMING=22.00
=
