tca tttgcctcgt gggattgggc
961 ttggtggtct tcttcttcag ttttctcctt tcaatatttc gttcaaagta ccatggctat
1021 ccatatag
Primer3 results:
No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 16 19 59.92 52.63 2.00 0.00 gattgggcttggtggtctt
RIGHT PRIMER 80 21 59.99 42.86 6.00 2.00 gccatggtactttgaacgaaa
SEQUENCE SIZE: 91
INCLUDED REGION SIZE: 91
PRODUCT SIZE: 65, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 2.00
1 tcatttgcctcgtgggattgggcttggtggtcttcttcttcagttttctcctttcaatat
>>>>>>>>>>>>>>>>>>> <
61 ttcgttcaaagtaccatggctatccatatag
<<<<<<<<<<<<<<<<<<<<
KEYS (in order of precedence):
>>>>>> left primer
<<<<<< right primer
ADDITIONAL OLIGOS
start len tm gc% any 3' seq
1 LEFT PRIMER 16 19 59.92 52.63 2.00 0.00 gattgggcttggtggtctt
RIGHT PRIMER 81 21 59.62 42.86 6.00 3.00 agccatggtactttgaacgaa
PRODUCT SIZE: 66, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
2 LEFT PRIMER 17 19 59.92 52.63 2.00 0.00 attgggcttggtggtcttc
RIGHT PRIMER 81 21 59.62 42.86 6.00 3.00 agccatggtactttgaacgaa
PRODUCT SIZE: 65, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 3.00
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 81 0 0 0 0 0 18 53 0 0 1 0 9
Right 81 0 0 0 0 0 79 0 0 0 0 0 2
Pair Stats:
considered 18, unacceptable product size 14, tm diff too large 1, ok 3
primer3 release 1.0
Check primers in blast exact:
LEFT PRIMER 16 19 59.92 52.63 2.00 0.00 gattgggcttggtggtctt
RIGHT PRIMER 80 21 59.99 42.86 6.00 2.00 gccatggtactttgaacgaaa
Primers ok in blast exact search.
Exon used in Primer 3
Primer 3 Sequence: 137-309
agaa ccttttggct gaaaaagtgg agcagctgat ggaatggagt
181 tccaggcgct cgatcttcag aatgaacggc gacaagttcc ggaagtttgt aaaagcccca
241 cctcggaact actccatgat cgtcatgttc actgctctgc agcctcagcg gcagtgttct
301 gtgtgcagg
Primer 3 Results:
No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 19 20 60.94 50.00 6.00 2.00 aagtggagcagctgatggaa
RIGHT PRIMER 83 20 60.65 50.00 4.00 1.00 gaacttgtcgccgttcattc
SEQUENCE SIZE: 173
INCLUDED REGION SIZE: 173
PRODUCT SIZE: 65, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 3.00
1 agaaccttttggctgaaaaagtggagcagctgatggaatggagttccaggcgctcgatct
>>>>>>>>>>>>>>>>>>>>
61 tcagaatgaacggcgacaagttccggaagtttgtaaaagccccacctcggaactactcca
<<<<<<<<<<<<<<<<<<<<
121 tgatcgtcatgttcactgctctgcagcctcagcggcagtgttctgtgtgcagg
KEYS (in order of precedence):
>>>>>> left primer
<<<<<< right primer
ADDITIONAL OLIGOS
start len tm gc% any 3' seq
1 LEFT PRIMER 18 20 60.94 50.00 6.00 4.00 aaagtggagcagctgatgga
RIGHT PRIMER 83 20 60.65 50.00 4.00 1.00 gaacttgtcgccgttcattc
PRODUCT SIZE: 66, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
2 LEFT PRIMER 51 21 59.97 47.62 5.00 5.00 cgctcgatcttcagaatgaac
RIGHT PRIMER 123 21 60.12 52.38 4.00 0.00 tcatggagtagttccgaggtg
PRODUCT SIZE: 73, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 3.00
3 LEFT PRIMER 50 19 59.64 52.63 5.00 3.00 gcgctcgatcttcagaatg
RIGHT PRIMER 123 21 60.12 52.38 4.00 0.00 tcatggagtagttccgaggtg
PRODUCT SIZE: 74, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
4 LEFT PRIMER 49 19 60.46 52.63 5.00 3.00 ggcgctcgatcttcagaat
RIGHT PRIMER 123 21 60.12 52.38 4.00 0.00 tcatggagtagttccgaggtg
PRODUCT SIZE: 75, PAIR ANY COMPL: 4.00, 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 243 0 0 0 0 0 56 111 3 0 42 7 24
Right 274 0 0 0 0 0 94 127 2 0 23 10 18
Pair Stats:
considered 341, unacceptable product size 332, tm diff too large 4, ok 5
primer3 release 1.0
Check Primers in Blast exact search:
LEFT PRIMER 19 20 60.94 50.00 6.00 2.00 aagtggagcagctgatggaa
RIGHT PRIMER 83 20 60.65 50.00 4.00 1.00 gaacttgtcgccgttcattc
Primers OK in Blast exact search!
VALIDATED PRIMER SEQUENCES:
FORWARD PRIMER: AAGTGGAGCAGCTGATGGAA
REVERSE PRIMER: GAACTTGTCGCCGTTCATTC