WARNING: Numbers in input sequence were deleted.
PRIMER PICKING RESULTS FOR gt01 m15
No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 47 20 60.60 55.00 3.00 1.00 gccctgcttcttgggatact
RIGHT PRIMER 130 20 59.83 60.00 4.00 2.00 cagggatagcgtctccactc
SEQUENCE SIZE: 232
INCLUDED REGION SIZE: 232
PRODUCT SIZE: 84, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
1 ggtgctgtgtaatggaccaggaacatgcgtccctatctgtgtgtccgccctgcttcttgg
>>>>>>>>>>>>>>
61 gatacttggagtgaagaaagtgatcatcgtctatgtcgagagcatctgccgagtggagac
>>>>>> <<<<<<<<<<
121 gctatccctgtcagggaagatcctgaggcacctctccgactacttcattgttcagtggcc
<<<<<<<<<<
181 cactctcaaggagaagtaccccaagtctgtgtacctggggcgaattgtttga
KEYS (in order of precedence):
>>>>>> left primer
<<<<<< right primer
ADDITIONAL OLIGOS
start len tm gc% any 3' seq
1 LEFT PRIMER 47 19 59.64 57.89 3.00 2.00 gccctgcttcttgggatac
RIGHT PRIMER 130 20 59.83 60.00 4.00 2.00 cagggatagcgtctccactc
PRODUCT SIZE: 84, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00
2 LEFT PRIMER 24 20 60.14 55.00 4.00 0.00 catgcgtccctatctgtgtg
RIGHT PRIMER 104 21 59.56 47.62 5.00 4.00 tgctctcgacatagacgatga
PRODUCT SIZE: 81, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 4.00
3 LEFT PRIMER 26 20 59.71 55.00 2.00 1.00 tgcgtccctatctgtgtgtc
RIGHT PRIMER 104 21 59.56 47.62 5.00 4.00 tgctctcgacatagacgatga
PRODUCT SIZE: 79, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 4.00
4 LEFT PRIMER 26 20 59.71 55.00 2.00 1.00 tgcgtccctatctgtgtgtc
RIGHT PRIMER 110 21 60.52 52.38 4.00 2.00 ggcagatgctctcgacataga
PRODUCT SIZE: 85, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 4.00
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 458 0 0 0 0 0 227 173 1 0 0 10 47
Right 397 0 0 0 0 0 197 120 5 0 32 13 30
Pair Stats:
considered 533, unacceptable product size 523, ok 10
primer3 release 1.0
(primer3_www_results.cgi v 0.4)
Primers to test for Exon 1 (423-654): Both pairs OK in Blast search
LEFT PRIMER 47 20 60.60 55.00 3.00 1.00 gccctgcttcttgggatact
RIGHT PRIMER 130 20 59.83 60.00 4.00 2.00 cagggatagcgtctccactc
SEQUENCE SIZE: 232
INCLUDED REGION SIZE: 232
PRODUCT SIZE: 84, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
2 LEFT PRIMER 24 20 60.14 55.00 4.00 0.00 catgcgtccctatctgtgtg
RIGHT PRIMER 104 21 59.56 47.62 5.00 4.00 tgctctcgacatagacgatga
PRODUCT SIZE: 81, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 4.00
Exon 2 (136-291)
ggtgg acacaccact gagatcttga ggctggttgg aagcttgtcc
181 aatgcctatt caccaaggca ttatgtcatt gctgagtctg atgaaatgag tgccaagaaa
241 atccattctc ttgaagaact ctctcgagcc cagaatgact ctactactga a
Primer3 Report for 136-291
PRIMER PICKING RESULTS FOR GT1M15
No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 1 21 60.44 57.14 5.00 4.00 ggtggacacaccactgagatc
RIGHT PRIMER 65 20 60.47 45.00 5.00 3.00 tgccttggtgaataggcatt
SEQUENCE SIZE: 156
INCLUDED REGION SIZE: 156
PRODUCT SIZE: 65, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
1 ggtggacacaccactgagatcttgaggctggttggaagcttgtccaatgcctattcacca
>>>>>>>>>>>>>>>>>>>>> <<<<<<<<<<<<<<<
61 aggcattatgtcattgctgagtctgatgaaatgagtgccaagaaaatccattctcttgaa
<<<<<
121 gaactctctcgagcccagaatgactctactactgaa
KEYS (in order of precedence):
>>>>>> left primer
<<<<<< right primer
ADDITIONAL OLIGOS
start len tm gc% any 3' seq
1 LEFT PRIMER 34 20 60.21 50.00 6.00 2.00 ggaagcttgtccaatgccta
RIGHT PRIMER 104 21 59.26 38.10 3.00 2.00 ttcttggcactcatttcatca
PRODUCT SIZE: 71, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
2 LEFT PRIMER 32 19 59.79 47.37 6.00 4.00 ttggaagcttgtccaatgc
RIGHT PRIMER 104 21 59.26 38.10 3.00 2.00 ttcttggcactcatttcatca
PRODUCT SIZE: 73, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
3 LEFT PRIMER 37 21 59.72 42.86 4.00 2.00 agcttgtccaatgcctattca
RIGHT PRIMER 104 21 59.26 38.10 3.00 2.00 ttcttggcactcatttcatca
PRODUCT SIZE: 68, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
4 LEFT PRIMER 1 21 60.44 57.14 5.00 4.00 ggtggacacaccactgagatc
RIGHT PRIMER 66 21 60.71 42.86 6.00 3.00 atgccttggtgaataggcatt
PRODUCT SIZE: 66, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
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 258 0 0 0 0 0 176 33 0 0 7 7 35
Right 239 0 0 0 0 0 187 8 3 1 18 2 20
Pair Stats:
considered 739, unacceptable product size 731, tm diff too large 2, ok 6
primer3 release 1.0
Primers to test for Exon 2 (136-291): Both pairs OK in Blast search
LEFT PRIMER 1 21 60.44 57.14 5.00 4.00 ggtggacacaccactgagatc
RIGHT PRIMER 65 20 60.47 45.00 5.00 3.00 tgccttggtgaataggcatt
SEQUENCE SIZE: 156
INCLUDED REGION SIZE: 156
PRODUCT SIZE: 65, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
3 LEFT PRIMER 37 21 59.72 42.86 4.00 2.00 agcttgtccaatgcctattca
RIGHT PRIMER 104 21 59.26 38.10 3.00 2.00 ttcttggcactcatttcatca
PRODUCT SIZE: 68, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
Sequence 1: lcl|seq_1
Length = 40 (1 .. 40)
Sequence 2: lcl|seq_2
Length = 654 (1 .. 654)
2
1
NOTE:Bitscore and expect value are calculated based on the size of the nr database.
NOTE:If protein translation is reversed, please repeat the search with reverse strand of the query sequence.
Score = 39.1 bits (20), Expect = 0.46
Identities = 20/20 (100%), Gaps = 0/20 (0%)
Strand=Plus/Plus
Query 1 GCCCTGCTTCTTGGGATACT 20
||||||||||||||||||||
Sbjct 469 GCCCTGCTTCTTGGGATACT 488
Score = 39.1 bits (20), Expect = 0.46
Identities = 20/20 (100%), Gaps = 0/20 (0%)
Strand=Plus/Minus
Query 21 CAGGGATAGCGTCTCCACTC 40
||||||||||||||||||||
Sbjct 552 CAGGGATAGCGTCTCCACTC 533
CPU time: 0.02 user secs. 0.01 sys. secs 0.03 total secs.
Lambda K H
1.33 0.621 1.12
Gapped
Lambda K H
1.33 0.621 1.12
Matrix: blastn matrix:1 -2
Gap Penalties: Existence: 5, Extension: 2
Number of Sequences: 1
Number of Hits to DB: 8
Number of extensions: 2
Number of successful extensions: 2
Number of sequences better than 10.0: 1
Number of HSP's gapped: 2
Number of HSP's successfully gapped: 2
Length of query: 40
Length of database: 16,556,997,203
Length adjustment: 23
Effective length of query: 17
Effective length of database: 16,556,997,180
Effective search space: 281468952060
Effective search space used: 281468952060
X1: 11 (21.1 bits)
X2: 26 (50.0 bits)
X3: 26 (50.0 bits)
S1: 11 (21.8 bits)
S2: 18 (35.3 bits)
Primers that passed in testing:
Forward: GCCCTGCTTCTTGGGATACT
Reverse: CAGGGATAGCGTCTCCACTC