PRIMER_DNTP_CONC=0.0
PRIMER_LIB_AMBIGUITY_CODES_CONSENSUS=1
PRIMER_THERMODYNAMIC_OLIGO_ALIGNMENT=1
PRIMER_TM_FORMULA=1
PRIMER_SALT_CORRECTIONS=1
PRIMER_SALT_DIVALENT=1.5
SEQUENCE_ID=anyway_1
PRIMER_EXPLAIN_FLAG=1
PRIMER_MAX_SELF_ANY=12
PRIMER_PAIR_MAX_COMPL_ANY=12
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=8
PRIMER_PAIR_MAX_COMPL_END=8
PRIMER_NUM_RETURN=1
PRIMER_PICK_ANYWAY=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_EXPLAIN=considered 10, low tm 7, high hairpin stability 3, ok 0
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_EXPLAIN=considered 0, ok 0
PRIMER_LEFT_NUM_RETURNED=0
PRIMER_RIGHT_NUM_RETURNED=0
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
=
SEQUENCE_ID=high_tm
PRIMER_MAX_SELF_ANY=12
PRIMER_PAIR_MAX_COMPL_ANY=12
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=8
PRIMER_PAIR_MAX_COMPL_END=8
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122
SEQUENCE_PRIMER=TCCTACAGCTGTGGGAAAATG
PRIMER_MAX_TM=60
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.436785
PRIMER_LEFT_0_PENALTY=2.141348
PRIMER_RIGHT_0_PENALTY=2.295437
PRIMER_LEFT_0_PROBLEMS= Hairpin stability too high;
PRIMER_LEFT_0_SEQUENCE=TCCTACAGCTGTGGGAAAATG
PRIMER_RIGHT_0_SEQUENCE=CAGTGTGTACATGGCGGC
PRIMER_LEFT_0=0,21
PRIMER_RIGHT_0=160,18
PRIMER_LEFT_0_TM=58.859
PRIMER_RIGHT_0_TM=59.705
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=61.111
PRIMER_LEFT_0_SELF_ANY_TH=41.37
PRIMER_RIGHT_0_SELF_ANY_TH=0.27
PRIMER_LEFT_0_SELF_END_TH=20.66
PRIMER_RIGHT_0_SELF_END_TH=0.27
PRIMER_LEFT_0_HAIRPIN_TH=50.77
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=2.3200
PRIMER_RIGHT_0_END_STABILITY=6.5300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=161
PRIMER_PAIR_0_PRODUCT_TM=84.1
=
SEQUENCE_ID=high_tm_and_self_any
PRIMER_MAX_SELF_ANY=11
PRIMER_PAIR_MAX_COMPL_ANY=11
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=8
PRIMER_PAIR_MAX_COMPL_END=8
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122
SEQUENCE_PRIMER=TCCTACAGCTGTGGGAAAATG
PRIMER_MAX_TM=60
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.436785
PRIMER_LEFT_0_PENALTY=2.141348
PRIMER_RIGHT_0_PENALTY=2.295437
PRIMER_LEFT_0_PROBLEMS= Hairpin stability too high;
PRIMER_LEFT_0_SEQUENCE=TCCTACAGCTGTGGGAAAATG
PRIMER_RIGHT_0_SEQUENCE=CAGTGTGTACATGGCGGC
PRIMER_LEFT_0=0,21
PRIMER_RIGHT_0=160,18
PRIMER_LEFT_0_TM=58.859
PRIMER_RIGHT_0_TM=59.705
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=61.111
PRIMER_LEFT_0_SELF_ANY_TH=41.37
PRIMER_RIGHT_0_SELF_ANY_TH=0.27
PRIMER_LEFT_0_SELF_END_TH=20.66
PRIMER_RIGHT_0_SELF_END_TH=0.27
PRIMER_LEFT_0_HAIRPIN_TH=50.77
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=2.3200
PRIMER_RIGHT_0_END_STABILITY=6.5300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=161
PRIMER_PAIR_0_PRODUCT_TM=84.1
=
SEQUENCE_ID=high_tm_and_self_any_and_target
PRIMER_MAX_SELF_ANY=11
PRIMER_PAIR_MAX_COMPL_ANY=11
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=8
PRIMER_PAIR_MAX_COMPL_END=8
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=20,122
SEQUENCE_PRIMER=TCCTACAGCTGTGGGAAAATG
PRIMER_MAX_TM=60
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_RIGHT_EXPLAIN=considered 3, high tm 2, ok 1
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.436785
PRIMER_LEFT_0_PENALTY=2.141348
PRIMER_RIGHT_0_PENALTY=2.295437
PRIMER_LEFT_0_PROBLEMS= Overlaps target; Hairpin stability too high;
PRIMER_LEFT_0_SEQUENCE=TCCTACAGCTGTGGGAAAATG
PRIMER_RIGHT_0_SEQUENCE=CAGTGTGTACATGGCGGC
PRIMER_LEFT_0=0,21
PRIMER_RIGHT_0=160,18
PRIMER_LEFT_0_TM=58.859
PRIMER_RIGHT_0_TM=59.705
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=61.111
PRIMER_LEFT_0_SELF_ANY_TH=41.37
PRIMER_RIGHT_0_SELF_ANY_TH=0.27
PRIMER_LEFT_0_SELF_END_TH=20.66
PRIMER_RIGHT_0_SELF_END_TH=0.27
PRIMER_LEFT_0_HAIRPIN_TH=50.77
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=2.3200
PRIMER_RIGHT_0_END_STABILITY=6.5300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=161
PRIMER_PAIR_0_PRODUCT_TM=84.1
=
SEQUENCE_ID=high_tm_and_self_any_and_excl_region
PRIMER_MAX_SELF_ANY=11
PRIMER_PAIR_MAX_COMPL_ANY=11
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=8
PRIMER_PAIR_MAX_COMPL_END=8
SEQUENCE_EXCLUDED_REGION=20,1
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122
SEQUENCE_PRIMER=TCCTACAGCTGTGGGAAAATG
PRIMER_MAX_TM=60
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.436785
PRIMER_LEFT_0_PENALTY=2.141348
PRIMER_RIGHT_0_PENALTY=2.295437
PRIMER_LEFT_0_PROBLEMS= Overlaps an excluded region; Hairpin stability too high;
PRIMER_LEFT_0_SEQUENCE=TCCTACAGCTGTGGGAAAATG
PRIMER_RIGHT_0_SEQUENCE=CAGTGTGTACATGGCGGC
PRIMER_LEFT_0=0,21
PRIMER_RIGHT_0=160,18
PRIMER_LEFT_0_TM=58.859
PRIMER_RIGHT_0_TM=59.705
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=61.111
PRIMER_LEFT_0_SELF_ANY_TH=41.37
PRIMER_RIGHT_0_SELF_ANY_TH=0.27
PRIMER_LEFT_0_SELF_END_TH=20.66
PRIMER_RIGHT_0_SELF_END_TH=0.27
PRIMER_LEFT_0_HAIRPIN_TH=50.77
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=2.3200
PRIMER_RIGHT_0_END_STABILITY=6.5300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=161
PRIMER_PAIR_0_PRODUCT_TM=84.1
=
SEQUENCE_ID=high_tm_and_self_any_and_incl_region
PRIMER_MAX_SELF_ANY=11
PRIMER_PAIR_MAX_COMPL_ANY=11
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=8
PRIMER_PAIR_MAX_COMPL_END=8
SEQUENCE_INCLUDED_REGION=1,160
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122
SEQUENCE_PRIMER=TCCTACAGCTGTGGGAAAATG
PRIMER_MAX_TM=60
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_ERROR=Specified left primer not in Included Region
=
SEQUENCE_ID=left_high_tm_and_self_any_and_right
PRIMER_MAX_SELF_ANY=11
PRIMER_PAIR_MAX_COMPL_ANY=11
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=8
PRIMER_PAIR_MAX_COMPL_END=8
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122
SEQUENCE_PRIMER=TCCTACAGCTGTGGGAAAATG
SEQUENCE_PRIMER_REVCOMP=CAGTGTGTACATGGCGGC
PRIMER_MAX_TM=60
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.436785
PRIMER_LEFT_0_PENALTY=2.141348
PRIMER_RIGHT_0_PENALTY=2.295437
PRIMER_LEFT_0_PROBLEMS= Hairpin stability too high;
PRIMER_LEFT_0_SEQUENCE=TCCTACAGCTGTGGGAAAATG
PRIMER_RIGHT_0_SEQUENCE=CAGTGTGTACATGGCGGC
PRIMER_LEFT_0=0,21
PRIMER_RIGHT_0=160,18
PRIMER_LEFT_0_TM=58.859
PRIMER_RIGHT_0_TM=59.705
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=61.111
PRIMER_LEFT_0_SELF_ANY_TH=41.37
PRIMER_RIGHT_0_SELF_ANY_TH=0.27
PRIMER_LEFT_0_SELF_END_TH=20.66
PRIMER_RIGHT_0_SELF_END_TH=0.27
PRIMER_LEFT_0_HAIRPIN_TH=50.77
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=2.3200
PRIMER_RIGHT_0_END_STABILITY=6.5300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=161
PRIMER_PAIR_0_PRODUCT_TM=84.1
=
SEQUENCE_ID=left_high_tm_and_self_any_and_right_bad_gc
PRIMER_MAX_SELF_ANY=11
PRIMER_PAIR_MAX_COMPL_ANY=11
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=8
PRIMER_PAIR_MAX_COMPL_END=8
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122
SEQUENCE_PRIMER=TCCTACAGCTGTGGGAAAATG
SEQUENCE_PRIMER_REVCOMP=CAGTGTGTACATGGCGGC
PRIMER_MAX_TM=60
PRIMER_MAX_GC=60
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.436785
PRIMER_LEFT_0_PENALTY=2.141348
PRIMER_RIGHT_0_PENALTY=2.295437
PRIMER_LEFT_0_PROBLEMS= Hairpin stability too high;
PRIMER_RIGHT_0_PROBLEMS= GC content too high;
PRIMER_LEFT_0_SEQUENCE=TCCTACAGCTGTGGGAAAATG
PRIMER_RIGHT_0_SEQUENCE=CAGTGTGTACATGGCGGC
PRIMER_LEFT_0=0,21
PRIMER_RIGHT_0=160,18
PRIMER_LEFT_0_TM=58.859
PRIMER_RIGHT_0_TM=59.705
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=61.111
PRIMER_LEFT_0_SELF_ANY_TH=41.37
PRIMER_RIGHT_0_SELF_ANY_TH=0.27
PRIMER_LEFT_0_SELF_END_TH=20.66
PRIMER_RIGHT_0_SELF_END_TH=0.27
PRIMER_LEFT_0_HAIRPIN_TH=50.77
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=2.3200
PRIMER_RIGHT_0_END_STABILITY=6.5300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=161
PRIMER_PAIR_0_PRODUCT_TM=84.1
=
SEQUENCE_ID=left_high_tm_and_self_any_and_right_bad_gc_and_target
PRIMER_MAX_SELF_ANY=11
PRIMER_PAIR_MAX_COMPL_ANY=11
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=8
PRIMER_PAIR_MAX_COMPL_END=8
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122 143,1
SEQUENCE_PRIMER=TCCTACAGCTGTGGGAAAATG
SEQUENCE_PRIMER_REVCOMP=CAGTGTGTACATGGCGGC
PRIMER_MAX_TM=60
PRIMER_MAX_GC=60
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.436785
PRIMER_LEFT_0_PENALTY=2.141348
PRIMER_RIGHT_0_PENALTY=2.295437
PRIMER_LEFT_0_PROBLEMS= Hairpin stability too high;
PRIMER_RIGHT_0_PROBLEMS= Overlaps target; GC content too high;
PRIMER_LEFT_0_SEQUENCE=TCCTACAGCTGTGGGAAAATG
PRIMER_RIGHT_0_SEQUENCE=CAGTGTGTACATGGCGGC
PRIMER_LEFT_0=0,21
PRIMER_RIGHT_0=160,18
PRIMER_LEFT_0_TM=58.859
PRIMER_RIGHT_0_TM=59.705
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=61.111
PRIMER_LEFT_0_SELF_ANY_TH=41.37
PRIMER_RIGHT_0_SELF_ANY_TH=0.27
PRIMER_LEFT_0_SELF_END_TH=20.66
PRIMER_RIGHT_0_SELF_END_TH=0.27
PRIMER_LEFT_0_HAIRPIN_TH=50.77
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=2.3200
PRIMER_RIGHT_0_END_STABILITY=6.5300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=161
PRIMER_PAIR_0_PRODUCT_TM=84.1
=
SEQUENCE_ID=internal_too_short
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_MAX_SELF_ANY=11
PRIMER_PAIR_MAX_COMPL_ANY=11
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=8
PRIMER_PAIR_MAX_COMPL_END=8
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122 143,1
SEQUENCE_PRIMER=TCCTACAGCTGTGGGAAAATG
SEQUENCE_PRIMER_REVCOMP=CAGTGTGTACATGGCGGC
SEQUENCE_INTERNAL_OLIGO=gcgtacgatccat
PRIMER_MAX_TM=60
PRIMER_MAX_GC=60
PRIMER_WARNING=Specified internal oligo < PRIMER_INTERNAL_MIN_SIZE; PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=1
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.436785
PRIMER_LEFT_0_PENALTY=2.141348
PRIMER_RIGHT_0_PENALTY=2.295437
PRIMER_INTERNAL_0_PENALTY=28.729043
PRIMER_LEFT_0_PROBLEMS= Hairpin stability too high;
PRIMER_RIGHT_0_PROBLEMS= Overlaps target; GC content too high;
PRIMER_INTERNAL_0_PROBLEMS= Temperature too low; Too short;
PRIMER_LEFT_0_SEQUENCE=TCCTACAGCTGTGGGAAAATG
PRIMER_RIGHT_0_SEQUENCE=CAGTGTGTACATGGCGGC
PRIMER_INTERNAL_0_SEQUENCE=gcgtacgatccat
PRIMER_LEFT_0=0,21
PRIMER_RIGHT_0=160,18
PRIMER_INTERNAL_0=23,13
PRIMER_LEFT_0_TM=58.859
PRIMER_RIGHT_0_TM=59.705
PRIMER_INTERNAL_0_TM=38.271
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=61.111
PRIMER_INTERNAL_0_GC_PERCENT=53.846
PRIMER_INTERNAL_0_SELF_ANY_TH=18.49
PRIMER_LEFT_0_SELF_ANY_TH=41.37
PRIMER_RIGHT_0_SELF_ANY_TH=0.27
PRIMER_INTERNAL_0_SELF_END_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=20.66
PRIMER_RIGHT_0_SELF_END_TH=0.27
PRIMER_LEFT_0_HAIRPIN_TH=50.77
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_INTERNAL_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=2.3200
PRIMER_RIGHT_0_END_STABILITY=6.5300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=161
PRIMER_PAIR_0_PRODUCT_TM=84.1
=
SEQUENCE_ID=internal_ok
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_MAX_SELF_ANY=11
PRIMER_PAIR_MAX_COMPL_ANY=11
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=8
PRIMER_PAIR_MAX_COMPL_END=8
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122 143,1
SEQUENCE_PRIMER=TCCTACAGCTGTGGGAAAATG
SEQUENCE_PRIMER_REVCOMP=CAGTGTGTACATGGCGGC
SEQUENCE_INTERNAL_OLIGO=gcgtacgatccatgctagc
PRIMER_MAX_TM=60
PRIMER_MAX_GC=60
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=1
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.436785
PRIMER_LEFT_0_PENALTY=2.141348
PRIMER_RIGHT_0_PENALTY=2.295437
PRIMER_INTERNAL_0_PENALTY=7.995085
PRIMER_LEFT_0_PROBLEMS= Hairpin stability too high;
PRIMER_RIGHT_0_PROBLEMS= Overlaps target; GC content too high;
PRIMER_INTERNAL_0_PROBLEMS= Temperature too low;
PRIMER_LEFT_0_SEQUENCE=TCCTACAGCTGTGGGAAAATG
PRIMER_RIGHT_0_SEQUENCE=CAGTGTGTACATGGCGGC
PRIMER_INTERNAL_0_SEQUENCE=gcgtacgatccatgctagc
PRIMER_LEFT_0=0,21
PRIMER_RIGHT_0=160,18
PRIMER_INTERNAL_0=23,19
PRIMER_LEFT_0_TM=58.859
PRIMER_RIGHT_0_TM=59.705
PRIMER_INTERNAL_0_TM=53.005
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=61.111
PRIMER_INTERNAL_0_GC_PERCENT=57.895
PRIMER_INTERNAL_0_SELF_ANY_TH=18.49
PRIMER_LEFT_0_SELF_ANY_TH=41.37
PRIMER_RIGHT_0_SELF_ANY_TH=0.27
PRIMER_INTERNAL_0_SELF_END_TH=4.81
PRIMER_LEFT_0_SELF_END_TH=20.66
PRIMER_RIGHT_0_SELF_END_TH=0.27
PRIMER_LEFT_0_HAIRPIN_TH=50.77
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_INTERNAL_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=2.3200
PRIMER_RIGHT_0_END_STABILITY=6.5300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=161
PRIMER_PAIR_0_PRODUCT_TM=84.1
=
SEQUENCE_ID=internal_too_cold
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_MAX_SELF_ANY=11
PRIMER_PAIR_MAX_COMPL_ANY=11
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=8
PRIMER_PAIR_MAX_COMPL_END=8
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122 143,1
SEQUENCE_PRIMER=TCCTACAGCTGTGGGAAAATG
SEQUENCE_PRIMER_REVCOMP=CAGTGTGTACATGGCGGC
SEQUENCE_INTERNAL_OLIGO=gcgtacgatccatgctagc
PRIMER_INTERNAL_MIN_TM=60
PRIMER_MAX_TM=60
PRIMER_MAX_GC=60
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=1
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.436785
PRIMER_LEFT_0_PENALTY=2.141348
PRIMER_RIGHT_0_PENALTY=2.295437
PRIMER_INTERNAL_0_PENALTY=7.995085
PRIMER_LEFT_0_PROBLEMS= Hairpin stability too high;
PRIMER_RIGHT_0_PROBLEMS= Overlaps target; GC content too high;
PRIMER_INTERNAL_0_PROBLEMS= Temperature too low;
PRIMER_LEFT_0_SEQUENCE=TCCTACAGCTGTGGGAAAATG
PRIMER_RIGHT_0_SEQUENCE=CAGTGTGTACATGGCGGC
PRIMER_INTERNAL_0_SEQUENCE=gcgtacgatccatgctagc
PRIMER_LEFT_0=0,21
PRIMER_RIGHT_0=160,18
PRIMER_INTERNAL_0=23,19
PRIMER_LEFT_0_TM=58.859
PRIMER_RIGHT_0_TM=59.705
PRIMER_INTERNAL_0_TM=53.005
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=61.111
PRIMER_INTERNAL_0_GC_PERCENT=57.895
PRIMER_INTERNAL_0_SELF_ANY_TH=18.49
PRIMER_LEFT_0_SELF_ANY_TH=41.37
PRIMER_RIGHT_0_SELF_ANY_TH=0.27
PRIMER_INTERNAL_0_SELF_END_TH=4.81
PRIMER_LEFT_0_SELF_END_TH=20.66
PRIMER_RIGHT_0_SELF_END_TH=0.27
PRIMER_LEFT_0_HAIRPIN_TH=50.77
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_INTERNAL_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=2.3200
PRIMER_RIGHT_0_END_STABILITY=6.5300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=161
PRIMER_PAIR_0_PRODUCT_TM=84.1
=
SEQUENCE_ID=internal_too_cold2
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_MAX_SELF_ANY=11
PRIMER_PAIR_MAX_COMPL_ANY=11
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=12
PRIMER_PAIR_MAX_COMPL_END=12
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122
SEQUENCE_PRIMER=CCTACAGCTGTGGGAAAATG
SEQUENCE_INTERNAL_OLIGO=gcgtacgatccatgctagc
PRIMER_INTERNAL_MIN_TM=60
PRIMER_MAX_TM=100
PRIMER_MAX_GC=100
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=1
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.984574
PRIMER_LEFT_0_PENALTY=2.689137
PRIMER_RIGHT_0_PENALTY=2.295437
PRIMER_INTERNAL_0_PENALTY=7.995085
PRIMER_LEFT_0_PROBLEMS= Hairpin stability too high;
PRIMER_INTERNAL_0_PROBLEMS= Temperature too low;
PRIMER_LEFT_0_SEQUENCE=CCTACAGCTGTGGGAAAATG
PRIMER_RIGHT_0_SEQUENCE=CAGTGTGTACATGGCGGC
PRIMER_INTERNAL_0_SEQUENCE=gcgtacgatccatgctagc
PRIMER_LEFT_0=1,20
PRIMER_RIGHT_0=160,18
PRIMER_INTERNAL_0=23,19
PRIMER_LEFT_0_TM=57.311
PRIMER_RIGHT_0_TM=59.705
PRIMER_INTERNAL_0_TM=53.005
PRIMER_LEFT_0_GC_PERCENT=50.000
PRIMER_RIGHT_0_GC_PERCENT=61.111
PRIMER_INTERNAL_0_GC_PERCENT=57.895
PRIMER_INTERNAL_0_SELF_ANY_TH=18.49
PRIMER_LEFT_0_SELF_ANY_TH=39.31
PRIMER_RIGHT_0_SELF_ANY_TH=0.27
PRIMER_INTERNAL_0_SELF_END_TH=4.81
PRIMER_LEFT_0_SELF_END_TH=15.46
PRIMER_RIGHT_0_SELF_END_TH=0.27
PRIMER_LEFT_0_HAIRPIN_TH=48.92
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_INTERNAL_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=2.3200
PRIMER_RIGHT_0_END_STABILITY=6.5300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=160
PRIMER_PAIR_0_PRODUCT_TM=84.2
=
SEQUENCE_ID=mispriming
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_MAX_SELF_ANY=11
PRIMER_PAIR_MAX_COMPL_ANY=11
PRIMER_MAX_POLY_X=100
PRIMER_INTERNAL_MAX_POLY_X=100
PRIMER_MAX_SELF_END=12
PRIMER_PAIR_MAX_COMPL_END=12
PRIMER_NUM_RETURN=1
SEQUENCE_TEMPLATE=TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagctatcatctactatcatctactatctactacacatctactatcatctacacacacacacacactaggccgctatatcatctagcGCCGCCATGTACACACTG
SEQUENCE_TARGET=21,122
SEQUENCE_PRIMER=CCTACAGCTGTGGGAAAATG
SEQUENCE_INTERNAL_OLIGO=gcgtacgatccatgctagc
PRIMER_INTERNAL_MIN_TM=60
PRIMER_MAX_TM=100
PRIMER_MAX_GC=100
PRIMER_MISPRIMING_LIBRARY=not_for_production.lib
PRIMER_MAX_LIBRARY_MISPRIMING=8
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_RIGHT_EXPLAIN=considered 1, ok 1
PRIMER_PAIR_EXPLAIN=considered 1, ok 1
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=1
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=4.984574
PRIMER_LEFT_0_PENALTY=2.689137
PRIMER_RIGHT_0_PENALTY=2.295437
PRIMER_INTERNAL_0_PENALTY=7.995085
PRIMER_LEFT_0_PROBLEMS= Hairpin stability too high; Similarity to non-template sequence too high;
PRIMER_INTERNAL_0_PROBLEMS= Temperature too low;
PRIMER_LEFT_0_SEQUENCE=CCTACAGCTGTGGGAAAATG
PRIMER_RIGHT_0_SEQUENCE=CAGTGTGTACATGGCGGC
PRIMER_INTERNAL_0_SEQUENCE=gcgtacgatccatgctagc
PRIMER_LEFT_0=1,20
PRIMER_RIGHT_0=160,18
PRIMER_INTERNAL_0=23,19
PRIMER_LEFT_0_TM=57.311
PRIMER_RIGHT_0_TM=59.705
PRIMER_INTERNAL_0_TM=53.005
PRIMER_LEFT_0_GC_PERCENT=50.000
PRIMER_RIGHT_0_GC_PERCENT=61.111
PRIMER_INTERNAL_0_GC_PERCENT=57.895
PRIMER_INTERNAL_0_SELF_ANY_TH=18.49
PRIMER_LEFT_0_SELF_ANY_TH=39.31
PRIMER_RIGHT_0_SELF_ANY_TH=0.27
PRIMER_INTERNAL_0_SELF_END_TH=4.81
PRIMER_LEFT_0_SELF_END_TH=15.46
PRIMER_RIGHT_0_SELF_END_TH=0.27
PRIMER_LEFT_0_HAIRPIN_TH=48.92
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_INTERNAL_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_LIBRARY_MISPRIMING=9.00, L1 Human L1 interspersed repetitive sequence - full length copy
PRIMER_RIGHT_0_LIBRARY_MISPRIMING=8.00, L1 Human L1 interspersed repetitive sequence - full length copy
PRIMER_PAIR_0_LIBRARY_MISPRIMING=17.00, L1 Human L1 interspersed repetitive sequence - full length copy
PRIMER_LEFT_0_END_STABILITY=2.3200
PRIMER_RIGHT_0_END_STABILITY=6.5300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=160
PRIMER_PAIR_0_PRODUCT_TM=84.2
=
