1
PRIMER_SEQUENCE_ID=quality_test1
3
COMMENT=quality scores out of range
5
PRIMER_INTERNAL_OLIGO_MIN_TM=0
6
PRIMER_INTERNAL_OLIGO_MAX_TM=100
10
PRIMER_PRODUCT_SIZE_RANGE=50-70 70-90 90-110 110-500
11
SEQUENCE=ATCGTAGCTGATCGTAGCTGATCGTAGCTAGTCAGTCTACTACTGATCTAGCTGATCATCGTAGCTAGCTAGTCATCATCGTACTACGTAGCTACTAGTCATCTAGCTAGCTAGTCGATGC
13
PRIMER_QUALITY_RANGE_MAX=40
14
PRIMER_SEQUENCE_QUALITY=50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50
16
PRIMER_SEQUENCE_ID=quality_test2
17
COMMENT=quality scores out of range
18
SEQUENCE=ATCGTAGCTGATCGTAGCTGATCGTAGCTAGTCAGTCTACTACTGATCTAGCTGATCATCGTAGCTAGCTAGTCATCATCGTACTACGTAGCTACTAGTCATCTAGCTAGCTAGTCGATGC
19
PRIMER_SEQUENCE_QUALITY=50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50
21
PRIMER_QUALITY_RANGE_MIN=60
22
PRIMER_QUALITY_RANGE_MAX=65
24
PRIMER_SEQUENCE_ID=quality_test3
25
COMMENT=quality scores out of range
26
SEQUENCE=ATCGTAGCTGATCGTAGCTGATCGTAGCTAGTCAGTCTACTACTGATCTAGCTGATCATCGTAGCTAGCTAGTCATCATCGTACTACGTAGCTACTAGTCATCTAGCTAGCTAGTCGATGC
27
PRIMER_SEQUENCE_QUALITY=50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50
28
PRIMER_QUALITY_RANGE_MIN=0
30
PRIMER_QUALITY_RANGE_MAX=49
32
PRIMER_SEQUENCE_ID=quality_test4
33
COMMENT=Should return exactly one pair
34
SEQUENCE=ATCGTAGCTGATCGTAGCTGATCGTAGCTAGTCAGTCTACTACTGATCTAGCTGATCATCGTAGCTAGCTAGTCATCATCGTACTACGTAGCTACTAGTCATCTAGCTAGCTAGTCGATGC
36
PRIMER_QUALITY_RANGE_MAX=70
38
PRIMER_SEQUENCE_QUALITY=60 60 60 60 60 60 60 60 60 60 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 60 60 60 60 60 60 60 60 60 60
40
PRIMER_SEQUENCE_ID=quality_test5
41
COMMENT=Should return exactly one pair
42
PRIMER_PICK_INTERNAL_OLIGO=1
43
PRIMER_INTERNAL_OLIGO_MIN_QUALITY=55
44
PRIMER_INTERNAL_OLIGO_MIN_SIZE=12
45
SEQUENCE=ATCGTAGCTGATCGTAGCTGATCGTAGCTAGTCAGTCTACTACTGATCTAGCTGATCATCGTAGCTAGCTAGTCATCATCGTACTACGTAGCTACTAGTCATCTAGCTAGCTAGTCGATGC
46
PRIMER_SEQUENCE_QUALITY=60 60 60 60 60 60 60 60 60 60 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 55 55 55 55 55 55 55 55 55 55 55 55 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 60 60 60 60 60 60 60 60 60 60
48
PRIMER_SEQUENCE_ID=quality_test6
49
COMMENT=Should fail because of low quality for middle oligo
50
PRIMER_PICK_INTERNAL_OLIGO=1
51
PRIMER_INTERNAL_OLIGO_MIN_QUALITY=51
52
SEQUENCE=ATCGTAGCTGATCGTAGCTGATCGTAGCTAGTCAGTCTACTACTGATCTAGCTGATCATCGTAGCTAGCTAGTCATCATCGTACTACGTAGCTACTAGTCATCTAGCTAGCTAGTCGATGC
53
PRIMER_SEQUENCE_QUALITY=60 60 60 60 60 60 60 60 60 60 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 53 54 54 54 54 50 54 54 54 54 54 51 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 60 60 60 60 60 60 60 60 60 60
55
PRIMER_SEQUENCE_ID=quality_test7
56
COMMENT=Should fail as there are too few score values
57
PRIMER_PICK_INTERNAL_OLIGO=1
58
PRIMER_INTERNAL_OLIGO_MIN_QUALITY=20
59
SEQUENCE=ATCGTAGCTGATCGTAGCTGATCGTAGCTAGTCAGTCTACTACTGATCTAGCTGATCATCGTAGCTAGCTAGTCATCATCGTACTACGTAGCTACTAGTCATCTAGCTAGCTAGTCGATGC
60
PRIMER_SEQUENCE_QUALITY=50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50
62
PRIMER_SEQUENCE_ID=quality_test8
63
COMMENT=Should fail as there are too many score values
64
PRIMER_PICK_INTERNAL_OLIGO=1
65
PRIMER_INTERNAL_OLIGO_MIN_QUALITY=20
66
SEQUENCE=ATCGTAGCTGATCGTAGCTGATCGTAGCTAGTCAGTCTACTACTGATCTAGCTGATCATCGTAGCTAGCTAGTCATCATCGTACTACGTAGCTACTAGTCATCTAGCTAGCTAGTCGATGC
67
PRIMER_SEQUENCE_QUALITY= 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50
69
PRIMER_SEQUENCE_ID=quality_test9
70
COMMENT=Should fail because of low end quality
71
PRIMER_PICK_INTERNAL_OLIGO=1
72
PRIMER_INTERNAL_OLIGO_MIN_QUALITY=55
73
PRIMER_INTERNAL_OLIGO_MIN_SIZE=12
74
PRIMER_MIN_END_QUALITY=61
75
SEQUENCE=ATCGTAGCTGATCGTAGCTGATCGTAGCTAGTCAGTCTACTACTGATCTAGCTGATCATCGTAGCTAGCTAGTCATCATCGTACTACGTAGCTACTAGTCATCTAGCTAGCTAGTCGATGC
76
PRIMER_SEQUENCE_QUALITY=60 60 60 60 60 60 60 60 60 60 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 55 55 55 55 55 55 55 55 55 55 55 55 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 60 60 60 60 60 60 60 60 60 60
78
PRIMER_SEQUENCE_ID=quality_test10
79
COMMENT=Should return exactly one pair
80
PRIMER_PICK_INTERNAL_OLIGO=1
81
PRIMER_INTERNAL_OLIGO_MIN_QUALITY=55
82
PRIMER_INTERNAL_OLIGO_MIN_SIZE=12
83
PRIMER_MIN_END_QUALITY=61
84
SEQUENCE=ATCGTAGCTGATCGTAGCTGATCGTAGCTAGTCAGTCTACTACTGATCTAGCTGATCATCGTAGCTAGCTAGTCATCATCGTACTACGTAGCTACTAGTCATCTAGCTAGCTAGTCGATGC
85
PRIMER_SEQUENCE_QUALITY=60 60 60 60 60 61 61 61 61 61 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 55 55 55 55 55 55 55 55 55 55 55 55 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 61 61 61 61 61 60 60 60 60 60
87
PRIMER_SEQUENCE_ID=quality_test11
88
COMMENT=Should fail as high quality pairs are not in the included region
89
PRIMER_PICK_INTERNAL_OLIGO=1
90
PRIMER_INTERNAL_OLIGO_MIN_QUALITY=55
91
PRIMER_INTERNAL_OLIGO_MIN_SIZE=12
92
PRIMER_MIN_END_QUALITY=61
94
SEQUENCE=ATCGTAGCTGATCGTAGCTGATCGTAGCTAGTCAGTCTACTACTGATCTAGCTGATCATCGTAGCTAGCTAGTCATCATCGTACTACGTAGCTACTAGTCATCTAGCTAGCTAGTCGATGC
95
PRIMER_SEQUENCE_QUALITY=60 60 60 60 60 61 61 61 61 61 61 61 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 55 55 55 55 55 55 55 55 55 55 55 55 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 61 61 61 61 61 60 60 60 60 60
97
PRIMER_SEQUENCE_ID=quality_test12
98
COMMENT=Should return exactly one pair
99
PRIMER_PICK_INTERNAL_OLIGO=1
100
PRIMER_INTERNAL_OLIGO_MIN_QUALITY=55
101
PRIMER_INTERNAL_OLIGO_MIN_SIZE=12
102
PRIMER_MIN_END_QUALITY=61
103
INCLUDED_REGION=2,115
104
SEQUENCE=ATCGTAGCTGATCGTAGCTGATCGTAGCTAGTCAGTCTACTACTGATCTAGCTGATCATCGTAGCTAGCTAGTCATCATCGTACTACGTAGCTACTAGTCATCTAGCTAGCTAGTCGATGC
105
PRIMER_SEQUENCE_QUALITY=60 60 60 60 60 61 61 61 61 61 61 61 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 55 55 55 55 55 55 55 55 55 55 55 55 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 61 61 61 61 61 61 61 61 61 60 50 50 50 50