help:请各位高手帮忙看看我设计的SD大鼠caspase-3的引物
丁香园论坛
2500
我最近用一在线的引物设计软件设计了一对SD大鼠caspase-3 的引物,不知行不,请高手多多帮忙.
结果如下:
Primer3 Output
--------------------------------------------------------------------------------
WARNING: Numbers in input sequence were deleted.
No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 1342 20 58.96 50.00 3.00 0.00 gaatccacgagcagagtcaa
RIGHT PRIMER 1557 20 60.25 50.00 3.00 0.00 tagccttcaacaagccaacc
SEQUENCE SIZE: 2484
INCLUDED REGION SIZE: 2484
PRODUCT SIZE: 216, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 0.00
1 gggatcaaagcttagtgtcctgaggtgcggagcttggaacgcgaagaaaagtgaccatgg
61 acaacaacgaaacctccgtggattcaaaatccattaataattttgaaacaaagactatcc
121 atggaagcaagtcgatggactctggaatatatctggacagcagttacaaaatggattacc
181 ctgaaatgggcttgtgtataataattaataataagaacttccataaaagcactggaatgt
241 cagctcgcaatggtaccgatgtcgatgcagctaacctcagagagacattcatggccctga
301 aatacgaagtcaggaataaaaatgaccttactcgtgaagaaattatggaattgatggata
361 gtgtttctaaggaagatcacagcaaaaggagcagttttgtgtgtgtgattctaagtcatg
421 gagatgaaggagtaatttttggaacgaacggacctgtggacctgaaaaaactaactagtt
481 tcttcagaggcgactactgccggagtctgactggaaagccgaaactcttcatcattcagg
541 cctgccgaggtacagagctggactgcggtattgagacagacagtggaactgacgatgata
601 tggcatgccagaagataccagtggaggccgacttcctgtatgcttactctaccgcacccg
661 gttactattcctggagaaattcaagggacgggtcatggttcatccagtcactttgcgcca
721 tgctgaaactgtacgcgcacaagctggaattcatgcacatcctcactcgtgttaaccgga
781 aggtggccacggaatttgagtccttctccctggacgccactttccatgcaaagaaacaga
841 tcccgtgtattgtgtcaatgctcacaaaagaactgtacttttatcactaaaggaatgact
901 gggagtggggtaggggcatgtttctgttttggtttttttttttggtttttgttttttgtt
961 tttttttttttatttgaatgccaaatgagaaaactgtcagggagactttttttttcccct
1021 ctcatttaaatcaaatccgatgttccaggtcgtcattgaacaataccactgcctgcaatg
1081 cagccacaatacaatacctcagctttgatatcaggtagttgaaatgaaatttaattagga
1141 acaaataaatgtcaataatggtacaaccatcatagaagaaattggaaagtcacactttgg
1201 taattagagagatttggtgccactatgaatttgaaattacgaagcagtgatgatcttata
1261 ccccctcctcacatctagaatgtcactttagttttcgttaagcataggaacgataatgtc
1321 aagtggtatgtcttagaacttgaatccacgagcagagtcaaaggctggttttggaactga
>>>>>>>>>>>>>>>>>>>>
1381 tctgtggaaatgaagagttggagcactgtagcacacattatagctactggttttgtgaca
1441 gttgaccaggtcacatatttattatttaagacaaaagtcgtgttttattattggaacaaa
1501 aataactttatatgagaggaaaaccatgtgtgaacttggttggcttgttgaaggctacca
<<<<<<<<<<<<<<<<<<<<
1561 tccttcagtggtggacatgacgacagggtgctacgatccaccagcatttgtttcttactt
1621 attcttatgcaaatcagcctaattttacagaccagcaaaatatctaaagagcgattatgc
1681 agcagcctcaaattgttgactattagaatggaataaaggaattctacaggaacagaccat
1741 aataccagctgtcagtcagagcgtaaggaaaggagaggtggcatctcctgtgattggaga
1801 gaagatggtttgagccggagcagagccatgggcacgtcttcagaaaaggcctgctagttg
1861 gacccaccttgtgagcactggtctgtgtggatgatgtccaagttaccacatgagactgaa
1921 ccagatcagaagctcctgcaaaaaggctttcccagagtgctgagcatctcaagaaagttc
1981 actggactccatgtacagagtctgtagcatgctgacttttctgttgaagtttacaatgaa
2041 aggaaaatgatggtgatatttgtaccgtcactagaaatgaagggccctggcacacgggac
2101 ttggaaagcatccagcaataggccctgcagcccatgtggaggagctgcagactggagtcc
2161 acccctcctccccatcagggcgtctgtgctccaggcttccttaatcatggaggtgctgag
2221 tcatcgctggctctccctgtgggtccagcatttcccataagcctccttattgcacaaaga
2281 aaggaaggaactcacattataactgatagtaggttcttgtttagactggatttactgtta
2341 actcttttctaagtaaatacagcatgtggtggtattttaaaatgtgttcccatgtgacaa
2401 ttttataaaagtcagatatcttgcatttttatttcaaaacataaattcagatttaaaatt
2461 aaaaaaaaaaaaaaaaaaaaaaaa
KEYS (in order of precedence):
>>>>>> left primer
<<<<<< right primer
ADDITIONAL OLIGOS
start len tm gc% any 3' seq
1 LEFT PRIMER 1342 20 58.96 50.00 3.00 0.00 gaatccacgagcagagtcaa
RIGHT PRIMER 1550 20 59.73 45.00 3.00 1.00 caacaagccaaccaagttca
PRODUCT SIZE: 209, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 2.00
2 LEFT PRIMER 1342 20 58.96 50.00 3.00 0.00 gaatccacgagcagagtcaa
RIGHT PRIMER 1554 20 60.66 50.00 3.00 0.00 ccttcaacaagccaaccaag
PRODUCT SIZE: 213, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 0.00
3 LEFT PRIMER 1342 20 58.96 50.00 3.00 0.00 gaatccacgagcagagtcaa
RIGHT PRIMER 1546 20 59.19 45.00 3.00 0.00 aagccaaccaagttcacaca
PRODUCT SIZE: 205, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 2.00
4 LEFT PRIMER 1342 20 58.96 50.00 3.00 0.00 gaatccacgagcagagtcaa
RIGHT PRIMER 1548 20 59.19 45.00 3.00 0.00 acaagccaaccaagttcaca
PRODUCT SIZE: 207, PAIR ANY COMPL: 3.00, 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 9195 0 0 0 698 0 2454 2424 693 0 1719 92 1115
Right 9393 0 0 0 1124 0 2439 2270 32 0 1692 88 1748
Pair Stats:
considered 61917, unacceptable product size 61821, high any compl 88, ok 8
primer3 release 1.0
(primer3_www_results.cgi v 0.4)
有同学说,有个引物评分,请问师兄我这对引物有多少分?谢谢!!!!!!
结果如下:
Primer3 Output
--------------------------------------------------------------------------------
WARNING: Numbers in input sequence were deleted.
No mispriming library specified
Using 1-based sequence positions
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 1342 20 58.96 50.00 3.00 0.00 gaatccacgagcagagtcaa
RIGHT PRIMER 1557 20 60.25 50.00 3.00 0.00 tagccttcaacaagccaacc
SEQUENCE SIZE: 2484
INCLUDED REGION SIZE: 2484
PRODUCT SIZE: 216, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 0.00
1 gggatcaaagcttagtgtcctgaggtgcggagcttggaacgcgaagaaaagtgaccatgg
61 acaacaacgaaacctccgtggattcaaaatccattaataattttgaaacaaagactatcc
121 atggaagcaagtcgatggactctggaatatatctggacagcagttacaaaatggattacc
181 ctgaaatgggcttgtgtataataattaataataagaacttccataaaagcactggaatgt
241 cagctcgcaatggtaccgatgtcgatgcagctaacctcagagagacattcatggccctga
301 aatacgaagtcaggaataaaaatgaccttactcgtgaagaaattatggaattgatggata
361 gtgtttctaaggaagatcacagcaaaaggagcagttttgtgtgtgtgattctaagtcatg
421 gagatgaaggagtaatttttggaacgaacggacctgtggacctgaaaaaactaactagtt
481 tcttcagaggcgactactgccggagtctgactggaaagccgaaactcttcatcattcagg
541 cctgccgaggtacagagctggactgcggtattgagacagacagtggaactgacgatgata
601 tggcatgccagaagataccagtggaggccgacttcctgtatgcttactctaccgcacccg
661 gttactattcctggagaaattcaagggacgggtcatggttcatccagtcactttgcgcca
721 tgctgaaactgtacgcgcacaagctggaattcatgcacatcctcactcgtgttaaccgga
781 aggtggccacggaatttgagtccttctccctggacgccactttccatgcaaagaaacaga
841 tcccgtgtattgtgtcaatgctcacaaaagaactgtacttttatcactaaaggaatgact
901 gggagtggggtaggggcatgtttctgttttggtttttttttttggtttttgttttttgtt
961 tttttttttttatttgaatgccaaatgagaaaactgtcagggagactttttttttcccct
1021 ctcatttaaatcaaatccgatgttccaggtcgtcattgaacaataccactgcctgcaatg
1081 cagccacaatacaatacctcagctttgatatcaggtagttgaaatgaaatttaattagga
1141 acaaataaatgtcaataatggtacaaccatcatagaagaaattggaaagtcacactttgg
1201 taattagagagatttggtgccactatgaatttgaaattacgaagcagtgatgatcttata
1261 ccccctcctcacatctagaatgtcactttagttttcgttaagcataggaacgataatgtc
1321 aagtggtatgtcttagaacttgaatccacgagcagagtcaaaggctggttttggaactga
>>>>>>>>>>>>>>>>>>>>
1381 tctgtggaaatgaagagttggagcactgtagcacacattatagctactggttttgtgaca
1441 gttgaccaggtcacatatttattatttaagacaaaagtcgtgttttattattggaacaaa
1501 aataactttatatgagaggaaaaccatgtgtgaacttggttggcttgttgaaggctacca
<<<<<<<<<<<<<<<<<<<<
1561 tccttcagtggtggacatgacgacagggtgctacgatccaccagcatttgtttcttactt
1621 attcttatgcaaatcagcctaattttacagaccagcaaaatatctaaagagcgattatgc
1681 agcagcctcaaattgttgactattagaatggaataaaggaattctacaggaacagaccat
1741 aataccagctgtcagtcagagcgtaaggaaaggagaggtggcatctcctgtgattggaga
1801 gaagatggtttgagccggagcagagccatgggcacgtcttcagaaaaggcctgctagttg
1861 gacccaccttgtgagcactggtctgtgtggatgatgtccaagttaccacatgagactgaa
1921 ccagatcagaagctcctgcaaaaaggctttcccagagtgctgagcatctcaagaaagttc
1981 actggactccatgtacagagtctgtagcatgctgacttttctgttgaagtttacaatgaa
2041 aggaaaatgatggtgatatttgtaccgtcactagaaatgaagggccctggcacacgggac
2101 ttggaaagcatccagcaataggccctgcagcccatgtggaggagctgcagactggagtcc
2161 acccctcctccccatcagggcgtctgtgctccaggcttccttaatcatggaggtgctgag
2221 tcatcgctggctctccctgtgggtccagcatttcccataagcctccttattgcacaaaga
2281 aaggaaggaactcacattataactgatagtaggttcttgtttagactggatttactgtta
2341 actcttttctaagtaaatacagcatgtggtggtattttaaaatgtgttcccatgtgacaa
2401 ttttataaaagtcagatatcttgcatttttatttcaaaacataaattcagatttaaaatt
2461 aaaaaaaaaaaaaaaaaaaaaaaa
KEYS (in order of precedence):
>>>>>> left primer
<<<<<< right primer
ADDITIONAL OLIGOS
start len tm gc% any 3' seq
1 LEFT PRIMER 1342 20 58.96 50.00 3.00 0.00 gaatccacgagcagagtcaa
RIGHT PRIMER 1550 20 59.73 45.00 3.00 1.00 caacaagccaaccaagttca
PRODUCT SIZE: 209, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 2.00
2 LEFT PRIMER 1342 20 58.96 50.00 3.00 0.00 gaatccacgagcagagtcaa
RIGHT PRIMER 1554 20 60.66 50.00 3.00 0.00 ccttcaacaagccaaccaag
PRODUCT SIZE: 213, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 0.00
3 LEFT PRIMER 1342 20 58.96 50.00 3.00 0.00 gaatccacgagcagagtcaa
RIGHT PRIMER 1546 20 59.19 45.00 3.00 0.00 aagccaaccaagttcacaca
PRODUCT SIZE: 205, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 2.00
4 LEFT PRIMER 1342 20 58.96 50.00 3.00 0.00 gaatccacgagcagagtcaa
RIGHT PRIMER 1548 20 59.19 45.00 3.00 0.00 acaagccaaccaagttcaca
PRODUCT SIZE: 207, PAIR ANY COMPL: 3.00, 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 9195 0 0 0 698 0 2454 2424 693 0 1719 92 1115
Right 9393 0 0 0 1124 0 2439 2270 32 0 1692 88 1748
Pair Stats:
considered 61917, unacceptable product size 61821, high any compl 88, ok 8
primer3 release 1.0
(primer3_www_results.cgi v 0.4)
有同学说,有个引物评分,请问师兄我这对引物有多少分?谢谢!!!!!!
