Primer 3 Output
WARNING: Numbers in input sequence were deleted.
PRIMER PICKING RESULTS FOR MGT M34
No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 36 20 60.01 50.00 3.00 0.00 gccaggaagtggtgtttgtt
RIGHT PRIMER 113 21 59.83 47.62 5.00 2.00 tcttcctttcagaggtgtcca
SEQUENCE SIZE: 200
INCLUDED REGION SIZE: 200
PRODUCT SIZE: 78, PAIR ANY COMPL: 6.00, PAIR 3' COMPL: 1.00
1 ccagacgtttgttcgacttcataaccagaagactggccaggaagtggtgtttgttgctga
>>>>>>>>>>>>>>>>>>>>
61 gccagataacaagaatgtatataagtttgaactggacacctctgaaaggaagattgaatt
<<<<<<<<<<<<<<<<<<<<<
121 tgactctgcctctggcacctacacactctacttaatcatcggggatgccacattgaagaa
181 cccaatcctctggaacgtgg
KEYS (in order of precedence):
>>>>>> left primer
<<<<<< right primer
ADDITIONAL OLIGOS
start len tm gc% any 3' seq
1 LEFT PRIMER 38 20 59.19 50.00 2.00 2.00 caggaagtggtgtttgttgc
RIGHT PRIMER 113 21 59.83 47.62 5.00 2.00 tcttcctttcagaggtgtcca
PRODUCT SIZE: 76, PAIR ANY COMPL: 6.00, PAIR 3' COMPL: 1.00
2 LEFT PRIMER 52 21 59.48 42.86 6.00 4.00 tgttgctgagccagataacaa
RIGHT PRIMER 136 20 59.43 50.00 4.00 4.00 gccagaggcagagtcaaatt
PRODUCT SIZE: 85, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
3 LEFT PRIMER 37 20 59.03 50.00 3.00 0.00 ccaggaagtggtgtttgttg
RIGHT PRIMER 113 21 59.83 47.62 5.00 2.00 tcttcctttcagaggtgtcca
PRODUCT SIZE: 77, PAIR ANY COMPL: 6.00, PAIR 3' COMPL: 2.00
4 LEFT PRIMER 38 21 60.20 47.62 3.00 0.00 caggaagtggtgtttgttgct
RIGHT PRIMER 113 21 59.83 47.62 5.00 2.00 tcttcctttcagaggtgtcca
PRODUCT SIZE: 76, PAIR ANY COMPL: 6.00, PAIR 3' COMPL: 1.00
Statistics
con too in in no tm tm high high high
sid many tar excl bad GC too too any 3' poly end
ered Ns get reg GC% clamp low high compl compl X stab ok
Left 367 0 0 0 6 0 274 51 0 0 0 9 27
Right 333 0 0 0 6 0 251 35 0 0 18 6 17
Pair Stats:
considered 310, unacceptable product size 301, tm diff too large 2, high any compl 1, ok 6
primer3 release 1.0
(primer3_www_results.cgi v 0.4)
First pair of primers used: Failed
Sense gccaggaagtggtgtttgtt
Anti-sense tcttcctttcagaggtgtcca
2nd exon to test (1753-1883)
gcgatgct gggcctcatg tatatctact ggactcagct caacatgttc
1801 cagaccctca agtacctggc tgtcctgggc actgtgacgt ttctggctgg caaccgaatg
1861 ctggcccagc atgcagttaa gag
PRIMER PICKING RESULTS FOR MGT M34
No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 5 21 59.57 47.62 4.00 2.00 tgctgggcctcatgtatatct
RIGHT PRIMER 75 21 59.92 57.14 6.00 4.00 caggacagccaggtacttgag
SEQUENCE SIZE: 131
INCLUDED REGION SIZE: 131
PRODUCT SIZE: 71, PAIR ANY COMPL: 6.00, PAIR 3' COMPL: 3.00
1 gcgatgctgggcctcatgtatatctactggactcagctcaacatgttccagaccctcaag
>>>>>>>>>>>>>>>>>>>>> <<<<<<
61 tacctggctgtcctgggcactgtgacgtttctggctggcaaccgaatgctggcccagcat
<<<<<<<<<<<<<<<
121 gcagttaagag
KEYS (in order of precedence):
>>>>>> left primer
<<<<<< right primer
ADDITIONAL OLIGOS
start len tm gc% any 3' seq
1 LEFT PRIMER 5 21 59.57 47.62 4.00 2.00 tgctgggcctcatgtatatct
RIGHT PRIMER 77 19 60.12 63.16 4.00 2.00 cccaggacagccaggtact
PRODUCT SIZE: 73, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 2.00
Statistics
con too in in no tm tm high high high
sid many tar excl bad GC too too any 3' poly end
ered Ns get reg GC% clamp low high compl compl X stab ok
Left 194 0 0 0 0 0 135 36 2 0 0 8 13
Right 190 0 0 0 0 0 22 146 0 0 0 8 14
Pair Stats:
considered 193, unacceptable product size 184, tm diff too large 6, high any compl 1, ok 2
primer3 release 1.0
2nd Primer to test: FAILED
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 5 21 59.57 47.62 4.00 2.00 tgctgggcctcatgtatatct
RIGHT PRIMER 75 21 59.92 57.14 6.00 4.00 caggacagccaggtacttgag
SEQUENCE SIZE: 131
INCLUDED REGION SIZE: 131
3rd Primer to test: from exon #1296-1495: OK in Blast search FAILED
LEFT PRIMER 52 21 59.48 42.86 6.00 4.00 tgttgctgagccagataacaa
RIGHT PRIMER 136 20 59.43 50.00 4.00 4.00 gccagaggcagagtcaaatt
PRODUCT SIZE: 85, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
Sequence for primer design: 1753-1883
agccccct cgactgtgct gtcccagagc ctgtttaccc caaaacagga
1801 aattcagcac ctgttccgag agcctgagaa gaggcccccc acagtggtgt ccaatacatt
1861 cacggccctc atcctctcgc cct
Primer3 Report:
PRIMER PICKING RESULTS FOR MGT M34
No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 46 20 59.53 50.00 5.00 1.00 ggaaattcagcacctgttcc
RIGHT PRIMER 120 20 59.27 45.00 4.00 4.00 atgagggccgtgaatgtatt
SEQUENCE SIZE: 131
INCLUDED REGION SIZE: 131
PRODUCT SIZE: 75, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00
1 agccccctcgactgtgctgtcccagagcctgtttaccccaaaacaggaaattcagcacct
>>>>>>>>>>>>>>>
61 gttccgagagcctgagaagaggccccccacagtggtgtccaatacattcacggccctcat
>>>>> <<<<<<<<<<<<<<<<<<<<
121 cctctcgccct
KEYS (in order of precedence):
>>>>>> left primer
<<<<<< right primer
ADDITIONAL OLIGOS
start len tm gc% any 3' seq
1 LEFT PRIMER 52 19 60.12 57.89 4.00 1.00 tcagcacctgttccgagag
RIGHT PRIMER 121 20 59.78 50.00 4.00 2.00 gatgagggccgtgaatgtat
PRODUCT SIZE: 70, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 0.00
2 LEFT PRIMER 51 19 60.54 52.63 4.00 2.00 ttcagcacctgttccgaga
RIGHT PRIMER 121 20 59.78 50.00 4.00 2.00 gatgagggccgtgaatgtat
PRODUCT SIZE: 71, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
3 LEFT PRIMER 52 19 60.12 57.89 4.00 1.00 tcagcacctgttccgagag
RIGHT PRIMER 120 20 59.27 45.00 4.00 4.00 atgagggccgtgaatgtatt
PRODUCT SIZE: 69, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 0.00
4 LEFT PRIMER 9 19 60.62 63.16 4.00 1.00 cgactgtgctgtcccagag
RIGHT PRIMER 80 20 59.68 55.00 4.00 2.00 tcttctcaggctctcggaac
PRODUCT SIZE: 72, PAIR ANY COMPL: 6.00, PAIR 3' COMPL: 5.00
Statistics
con too in in no tm tm high high high
sid many tar excl bad GC too too any 3' poly end
ered Ns get reg GC% clamp low high compl compl X stab ok
Left 143 0 0 0 0 0 23 61 0 0 30 6 23
Right 161 0 0 0 0 0 37 80 0 0 20 2 22
Pair Stats:
considered 243, unacceptable product size 225, tm diff too large 5, high any compl 7, high end compl 1, ok 5
primer3 release 1.0
4th primer to test: OK in Blast search FAILED
LEFT PRIMER 46 20 59.53 50.00 5.00 1.00 ggaaattcagcacctgttcc
RIGHT PRIMER 120 20 59.27 45.00 4.00 4.00 atgagggccgtgaatgtatt
SEQUENCE SIZE: 131
INCLUDED REGION SIZE: 131
PRODUCT SIZE: 75, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00
3rd exon to test: 1184-1299
gggtgac ctacccagcc
1201 aaagccaagg gtacattcat cgcagacagc catcagaact ttgccctgtt tttccagctg
1261 gtagatgtga acaccggtgc ggagctcacc cctcaccag
PRIMER PICKING RESULTS FOR MGT M34
No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 20 20 59.96 50.00 4.00 2.00 agccaagggtacattcatcg
RIGHT PRIMER 94 20 59.01 55.00 4.00 1.00 ccggtgttcacatctaccag
SEQUENCE SIZE: 116
INCLUDED REGION SIZE: 116
PRODUCT SIZE: 75, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00
1 gggtgacctacccagccaaagccaagggtacattcatcgcagacagccatcagaactttg
>>>>>>>>>>>>>>>>>>>>
61 ccctgtttttccagctggtagatgtgaacaccggtgcggagctcacccctcaccag
<<<<<<<<<<<<<<<<<<<<
KEYS (in order of precedence):
>>>>>> left primer
<<<<<< right primer
ADDITIONAL OLIGOS
start len tm gc% any 3' seq
1 LEFT PRIMER 20 20 59.96 50.00 4.00 2.00 agccaagggtacattcatcg
RIGHT PRIMER 93 21 60.19 52.38 4.00 4.00 cggtgttcacatctaccagct
PRODUCT SIZE: 74, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
2 LEFT PRIMER 26 20 59.53 50.00 4.00 0.00 gggtacattcatcgcagaca
RIGHT PRIMER 94 20 59.01 55.00 4.00 1.00 ccggtgttcacatctaccag
PRODUCT SIZE: 69, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
3 LEFT PRIMER 26 20 59.53 50.00 4.00 0.00 gggtacattcatcgcagaca
RIGHT PRIMER 95 21 59.90 52.38 6.00 1.00 accggtgttcacatctaccag
PRODUCT SIZE: 70, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
4 LEFT PRIMER 26 20 59.53 50.00 4.00 0.00 gggtacattcatcgcagaca
RIGHT PRIMER 93 21 60.19 52.38 4.00 4.00 cggtgttcacatctaccagct
PRODUCT SIZE: 68, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00
Statistics
con too in in no tm tm high high high
sid many tar excl bad GC too too any 3' poly end
ered Ns get reg GC% clamp low high compl compl X stab ok
Left 138 0 0 0 0 0 39 68 0 0 4 7 20
Right 106 0 0 0 0 0 29 39 1 0 29 1 7
Pair Stats:
considered 55, unacceptable product size 43, tm diff too large 3, ok 9
primer3 release 1.0
4th exon to test: 1578-1677
tca gcacctgttc cgagagcctg agaagaggcc ccccacagtg
1621 gtgtccaata cattcacggc cctcatcctc tcgcccttgc tcctgctctt tgcactg
PRIMER PICKING RESULTS FOR MGT M34
No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 1 19 60.12 57.89 4.00 1.00 tcagcacctgttccgagag
RIGHT PRIMER 70 20 59.78 50.00 4.00 2.00 gatgagggccgtgaatgtat
SEQUENCE SIZE: 100
INCLUDED REGION SIZE: 100
PRODUCT SIZE: 70, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 0.00
1 tcagcacctgttccgagagcctgagaagaggccccccacagtggtgtccaatacattcac
>>>>>>>>>>>>>>>>>>> <<<<<<<<<<
61 ggccctcatcctctcgcccttgctcctgctctttgcactg
<<<<<<<<<<
KEYS (in order of precedence):
>>>>>> left primer
<<<<<< right primer
ADDITIONAL OLIGOS
start len tm gc% any 3' seq
1 LEFT PRIMER 1 19 60.12 57.89 4.00 1.00 tcagcacctgttccgagag
RIGHT PRIMER 69 20 59.27 45.00 4.00 4.00 atgagggccgtgaatgtatt
PRODUCT SIZE: 69, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 0.00
2 LEFT PRIMER 1 19 60.12 57.89 4.00 1.00 tcagcacctgttccgagag
RIGHT PRIMER 70 19 59.47 52.63 4.00 2.00 gatgagggccgtgaatgta
PRODUCT SIZE: 70, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 0.00
3 LEFT PRIMER 1 19 60.12 57.89 4.00 1.00 tcagcacctgttccgagag
RIGHT PRIMER 66 19 60.73 52.63 4.00 0.00 agggccgtgaatgtattgg
PRODUCT SIZE: 66, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 1.00
Statistics
con too in in no tm tm high high high
sid many tar excl bad GC too too any 3' poly end
ered Ns get reg GC% clamp low high compl compl X stab ok
Left 67 0 0 0 0 0 8 27 0 0 20 2 10
Right 108 0 0 0 0 0 3 97 0 0 0 1 7
Pair Stats:
considered 70, unacceptable product size 64, tm diff too large 2, ok 4
primer3 release 1.0
5th primer to test: OK in Blast search “a” GOOD
LEFT PRIMER 20 20 59.96 50.00 4.00 2.00 agccaagggtacattcatcg
RIGHT PRIMER 94 20 59.01 55.00 4.00 1.00 ccggtgttcacatctaccag
SEQUENCE SIZE: 116
INCLUDED REGION SIZE: 116
PRODUCT SIZE: 75, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00
6th primer to test: OK in Blast search “b” FAILED
LEFT PRIMER 1 19 60.12 57.89 4.00 1.00 tcagcacctgttccgagag
RIGHT PRIMER 70 20 59.78 50.00 4.00 2.00 gatgagggccgtgaatgtat
SEQUENCE SIZE: 100
INCLUDED REGION SIZE: 100
PRODUCT SIZE: 70, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 0.00