1
PRIMER PICKING RESULTS FOR anyway_1
3
No mispriming library specified
4
Using 0-based sequence positions
5
OLIGO start len tm gc% any 3' seq
6
LEFT PRIMER 0 21 60.12 47.62 12.00 0.00 TCCTACAGCTGTGGGAAAATG
7
RIGHT PRIMER 160 18 59.69 61.11 6.00 3.00 CAGTGTGTACATGGCGGC
9
INCLUDED REGION SIZE: 161
11
PRODUCT SIZE: 161, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
12
TARGETS (start, len)*: 21,122
14
0 TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagc
15
>>>>>>>>>>>>>>>>>>>>>***************************************
17
60 tatcatctactatcatctactatctactacacatctactatcatctacacacacacacac
18
************************************************************
20
120 actaggccgctatatcatctagcGCCGCCATGTACACACTG
21
***********************<<<<<<<<<<<<<<<<<<
23
KEYS (in order of precedence):
30
con too in in no tm tm high high high
31
sid many tar excl bad GC too too any 3' poly end
32
ered Ns get reg GC% clamp low high compl compl X stab ok
33
Left 10 0 0 0 0 0 7 0 0 0 0 0 3
34
Right 1 0 0 0 0 0 0 0 0 0 0 0 1
40
PRIMER PICKING RESULTS FOR high_tm
42
No mispriming library specified
43
Using 0-based sequence positions
44
WARNING: Left primer is unacceptable: Tm too high
46
OLIGO start len tm gc% any 3' seq
47
LEFT PRIMER 0 21 60.12 47.62 12.00 0.00 TCCTACAGCTGTGGGAAAATG
48
RIGHT PRIMER 160 18 59.69 61.11 6.00 3.00 CAGTGTGTACATGGCGGC
50
INCLUDED REGION SIZE: 161
52
PRODUCT SIZE: 161, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
53
TARGETS (start, len)*: 21,122
55
0 TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagc
56
>>>>>>>>>>>>>>>>>>>>>***************************************
58
60 tatcatctactatcatctactatctactacacatctactatcatctacacacacacacac
59
************************************************************
61
120 actaggccgctatatcatctagcGCCGCCATGTACACACTG
62
***********************<<<<<<<<<<<<<<<<<<
64
KEYS (in order of precedence):
71
con too in in no tm tm high high high
72
sid many tar excl bad GC too too any 3' poly end
73
ered Ns get reg GC% clamp low high compl compl X stab ok
74
Right 1 0 0 0 0 0 0 0 0 0 0 0 1
80
PRIMER PICKING RESULTS FOR high_tm_and_self_any
82
No mispriming library specified
83
Using 0-based sequence positions
84
WARNING: Left primer is unacceptable: Tm too high/High self complementarity
86
OLIGO start len tm gc% any 3' seq
87
LEFT PRIMER 0 21 60.12 47.62 12.00 0.00 TCCTACAGCTGTGGGAAAATG
88
RIGHT PRIMER 160 18 59.69 61.11 6.00 3.00 CAGTGTGTACATGGCGGC
90
INCLUDED REGION SIZE: 161
92
PRODUCT SIZE: 161, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
93
TARGETS (start, len)*: 21,122
95
0 TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagc
96
>>>>>>>>>>>>>>>>>>>>>***************************************
98
60 tatcatctactatcatctactatctactacacatctactatcatctacacacacacacac
99
************************************************************
101
120 actaggccgctatatcatctagcGCCGCCATGTACACACTG
102
***********************<<<<<<<<<<<<<<<<<<
104
KEYS (in order of precedence):
111
con too in in no tm tm high high high
112
sid many tar excl bad GC too too any 3' poly end
113
ered Ns get reg GC% clamp low high compl compl X stab ok
114
Right 1 0 0 0 0 0 0 0 0 0 0 0 1
120
PRIMER PICKING RESULTS FOR high_tm_and_self_any_and_target
122
No mispriming library specified
123
Using 0-based sequence positions
124
WARNING: Left primer is unacceptable: Overlaps Target/Tm too high/High self complementarity
126
OLIGO start len tm gc% any 3' seq
127
LEFT PRIMER 0 21 60.12 47.62 12.00 0.00 TCCTACAGCTGTGGGAAAATG
128
RIGHT PRIMER 160 18 59.69 61.11 6.00 3.00 CAGTGTGTACATGGCGGC
130
INCLUDED REGION SIZE: 161
132
PRODUCT SIZE: 161, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
133
TARGETS (start, len)*: 20,122
135
0 TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagc
136
>>>>>>>>>>>>>>>>>>>>)***************************************
138
60 tatcatctactatcatctactatctactacacatctactatcatctacacacacacacac
139
************************************************************
141
120 actaggccgctatatcatctagcGCCGCCATGTACACACTG
142
********************** <<<<<<<<<<<<<<<<<<
144
KEYS (in order of precedence):
151
con too in in no tm tm high high high
152
sid many tar excl bad GC too too any 3' poly end
153
ered Ns get reg GC% clamp low high compl compl X stab ok
154
Right 3 0 0 0 0 0 0 2 0 0 0 0 1
160
PRIMER PICKING RESULTS FOR high_tm_and_self_any_and_excl_region
162
No mispriming library specified
163
Using 0-based sequence positions
164
WARNING: Left primer is unacceptable: Overlaps Excluded Region/Tm too high/High self complementarity
166
OLIGO start len tm gc% any 3' seq
167
LEFT PRIMER 0 21 60.12 47.62 12.00 0.00 TCCTACAGCTGTGGGAAAATG
168
RIGHT PRIMER 160 18 59.69 61.11 6.00 3.00 CAGTGTGTACATGGCGGC
170
INCLUDED REGION SIZE: 161
172
PRODUCT SIZE: 161, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
173
TARGETS (start, len)*: 21,122
174
EXCLUDED REGIONS (start, len)*: 20,1
176
0 TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagc
177
>>>>>>>>>>>>>>>>>>>>X***************************************
179
60 tatcatctactatcatctactatctactacacatctactatcatctacacacacacacac
180
************************************************************
182
120 actaggccgctatatcatctagcGCCGCCATGTACACACTG
183
***********************<<<<<<<<<<<<<<<<<<
185
KEYS (in order of precedence):
186
XXXXXX excluded region
193
con too in in no tm tm high high high
194
sid many tar excl bad GC too too any 3' poly end
195
ered Ns get reg GC% clamp low high compl compl X stab ok
196
Right 1 0 0 0 0 0 0 0 0 0 0 0 1
202
PRIMER PICKING RESULTS FOR high_tm_and_self_any_and_incl_region
204
INPUT PROBLEM: Specified left primer not in Included Region
206
PRIMER PICKING RESULTS FOR left_high_tm_and_self_any_and_right
208
No mispriming library specified
209
Using 0-based sequence positions
210
WARNING: Left primer is unacceptable: Tm too high/High self complementarity
212
OLIGO start len tm gc% any 3' seq
213
LEFT PRIMER 0 21 60.12 47.62 12.00 0.00 TCCTACAGCTGTGGGAAAATG
214
RIGHT PRIMER 160 18 59.69 61.11 6.00 3.00 CAGTGTGTACATGGCGGC
216
INCLUDED REGION SIZE: 161
218
PRODUCT SIZE: 161, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
219
TARGETS (start, len)*: 21,122
221
0 TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagc
222
>>>>>>>>>>>>>>>>>>>>>***************************************
224
60 tatcatctactatcatctactatctactacacatctactatcatctacacacacacacac
225
************************************************************
227
120 actaggccgctatatcatctagcGCCGCCATGTACACACTG
228
***********************<<<<<<<<<<<<<<<<<<
230
KEYS (in order of precedence):
242
PRIMER PICKING RESULTS FOR left_high_tm_and_self_any_and_right_bad_gc
244
No mispriming library specified
245
Using 0-based sequence positions
246
WARNING: Left primer is unacceptable: Tm too high/High self complementarity; Right primer is unacceptable: Unacceptable GC content
248
OLIGO start len tm gc% any 3' seq
249
LEFT PRIMER 0 21 60.12 47.62 12.00 0.00 TCCTACAGCTGTGGGAAAATG
250
RIGHT PRIMER 160 18 59.69 61.11 6.00 3.00 CAGTGTGTACATGGCGGC
252
INCLUDED REGION SIZE: 161
254
PRODUCT SIZE: 161, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
255
TARGETS (start, len)*: 21,122
257
0 TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagc
258
>>>>>>>>>>>>>>>>>>>>>***************************************
260
60 tatcatctactatcatctactatctactacacatctactatcatctacacacacacacac
261
************************************************************
263
120 actaggccgctatatcatctagcGCCGCCATGTACACACTG
264
***********************<<<<<<<<<<<<<<<<<<
266
KEYS (in order of precedence):
278
PRIMER PICKING RESULTS FOR left_high_tm_and_self_any_and_right_bad_gc_and_target
280
No mispriming library specified
281
Using 0-based sequence positions
282
WARNING: Left primer is unacceptable: Tm too high/High self complementarity; Right primer is unacceptable: Overlaps Target/Unacceptable GC content
284
OLIGO start len tm gc% any 3' seq
285
LEFT PRIMER 0 21 60.12 47.62 12.00 0.00 TCCTACAGCTGTGGGAAAATG
286
RIGHT PRIMER 160 18 59.69 61.11 6.00 3.00 CAGTGTGTACATGGCGGC
288
INCLUDED REGION SIZE: 161
290
PRODUCT SIZE: 161, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
291
TARGETS (start, len)*: 21,122 143,1
293
0 TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagc
294
>>>>>>>>>>>>>>>>>>>>>***************************************
296
60 tatcatctactatcatctactatctactacacatctactatcatctacacacacacacac
297
************************************************************
299
120 actaggccgctatatcatctagcGCCGCCATGTACACACTG
300
***********************(<<<<<<<<<<<<<<<<<
302
KEYS (in order of precedence):
314
PRIMER PICKING RESULTS FOR internal_too_short
316
INPUT PROBLEM: Specified internal oligo too short
318
PRIMER PICKING RESULTS FOR internal_ok
320
No mispriming library specified
321
No internal oligo mishyb library specified
322
Using 0-based sequence positions
323
WARNING: Left primer is unacceptable: Tm too high/High self complementarity; Right primer is unacceptable: Overlaps Target/Unacceptable GC content
325
OLIGO start len tm gc% any 3' seq
326
LEFT PRIMER 0 21 60.12 47.62 12.00 0.00 TCCTACAGCTGTGGGAAAATG
327
RIGHT PRIMER 160 18 59.69 61.11 6.00 3.00 CAGTGTGTACATGGCGGC
328
INTERNAL OLIGO 23 19 59.42 57.89 6.00 6.00 gcgtacgatccatgctagc
330
INCLUDED REGION SIZE: 161
332
PRODUCT SIZE: 161, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
333
TARGETS (start, len)*: 21,122 143,1
335
0 TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagc
336
>>>>>>>>>>>>>>>>>>>>>***************************************
338
60 tatcatctactatcatctactatctactacacatctactatcatctacacacacacacac
339
************************************************************
341
120 actaggccgctatatcatctagcGCCGCCATGTACACACTG
342
***********************(<<<<<<<<<<<<<<<<<
344
KEYS (in order of precedence):
348
^^^^^^ internal oligo
357
PRIMER PICKING RESULTS FOR internal_too_cold
359
No mispriming library specified
360
No internal oligo mishyb library specified
361
Using 0-based sequence positions
362
WARNING: Left primer is unacceptable: Tm too high/High self complementarity; Right primer is unacceptable: Overlaps Target/Unacceptable GC content; Hybridization probe is unacceptable: Tm too low
364
OLIGO start len tm gc% any 3' seq
365
LEFT PRIMER 0 21 60.12 47.62 12.00 0.00 TCCTACAGCTGTGGGAAAATG
366
RIGHT PRIMER 160 18 59.69 61.11 6.00 3.00 CAGTGTGTACATGGCGGC
367
INTERNAL OLIGO 23 19 59.42 57.89 6.00 6.00 gcgtacgatccatgctagc
369
INCLUDED REGION SIZE: 161
371
PRODUCT SIZE: 161, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
372
TARGETS (start, len)*: 21,122 143,1
374
0 TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagc
375
>>>>>>>>>>>>>>>>>>>>>***************************************
377
60 tatcatctactatcatctactatctactacacatctactatcatctacacacacacacac
378
************************************************************
380
120 actaggccgctatatcatctagcGCCGCCATGTACACACTG
381
***********************(<<<<<<<<<<<<<<<<<
383
KEYS (in order of precedence):
387
^^^^^^ internal oligo
396
PRIMER PICKING RESULTS FOR internal_too_cold2
398
No mispriming library specified
399
No internal oligo mishyb library specified
400
Using 0-based sequence positions
401
WARNING: Hybridization probe is unacceptable: Tm too low
403
OLIGO start len tm gc% any 3' seq
404
LEFT PRIMER 1 20 58.23 50.00 10.00 0.00 CCTACAGCTGTGGGAAAATG
405
RIGHT PRIMER 160 18 59.69 61.11 6.00 3.00 CAGTGTGTACATGGCGGC
406
INTERNAL OLIGO 23 19 59.42 57.89 6.00 6.00 gcgtacgatccatgctagc
408
INCLUDED REGION SIZE: 161
410
PRODUCT SIZE: 160, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
411
TARGETS (start, len)*: 21,122
413
0 TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagc
414
>>>>>>>>>>>>>>>>>>>>***************************************
416
60 tatcatctactatcatctactatctactacacatctactatcatctacacacacacacac
417
************************************************************
419
120 actaggccgctatatcatctagcGCCGCCATGTACACACTG
420
***********************<<<<<<<<<<<<<<<<<<
422
KEYS (in order of precedence):
426
^^^^^^ internal oligo
430
con too in in no tm tm high high high
431
sid many tar excl bad GC too too any 3' poly end
432
ered Ns get reg GC% clamp low high compl compl X stab ok
433
Right 1 0 0 0 0 0 0 0 0 0 0 0 1
439
PRIMER PICKING RESULTS FOR mispriming
441
Using mispriming library not_for_production.lib
442
No internal oligo mishyb library specified
443
Using 0-based sequence positions
444
WARNING: Left primer is unacceptable: High similarity to mispriming or mishyb library; Hybridization probe is unacceptable: Tm too low
446
OLIGO start len tm gc% any 3' rep seq
447
LEFT PRIMER 1 20 58.23 50.00 10.00 0.00 9.00 CCTACAGCTGTGGGAAAATG
448
RIGHT PRIMER 160 18 59.69 61.11 6.00 3.00 8.00 CAGTGTGTACATGGCGGC
449
INTERNAL OLIGO 23 19 59.42 57.89 6.00 6.00 gcgtacgatccatgctagc
451
INCLUDED REGION SIZE: 161
453
PRODUCT SIZE: 160, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
454
TARGETS (start, len)*: 21,122
456
0 TCCTACAGCTGTGGGAAAATGatgcgtacgatccatgctagctagctactatcgattagc
457
>>>>>>>>>>>>>>>>>>>>***************************************
459
60 tatcatctactatcatctactatctactacacatctactatcatctacacacacacacac
460
************************************************************
462
120 actaggccgctatatcatctagcGCCGCCATGTACACACTG
463
***********************<<<<<<<<<<<<<<<<<<
465
KEYS (in order of precedence):
469
^^^^^^ internal oligo
473
con too in in no tm tm high high high high
474
sid many tar excl bad GC too too any 3' lib poly end
475
ered Ns get reg GC% clamp low high compl compl sim X stab ok
476
Right 1 0 0 0 0 0 0 0 0 0 0 0 0 1
added
removed
test/primer_must_use_formatted_output
