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小师妹请教各位高手一下引物设计

丁香园论坛

638
各位尊敬的大师兄:
以下序列是人类某整段基因组DNA转录起始点上游2826BP长度片段,老板要我PCR出转录起始点上游1500BP片段,需要设计引物,我才开始做,希望各位高手赐教.小师妹不慎感激!!!

1 tgacagaagg agaccctgtc tcaaaaataa taagaataat aattaataat aataggccaa
61 accaaatacc catcaccttc tgctgtgcct cccctttccc caataaatcc agtgtcttgc
121 tttcaaattt tgtggttaaa aaagatgatg agtttctaag acgtgggggc taaagcttgt
181 ttggccgttt tagggtttgt tggaattttt ttttcgtcta tgtacttgtg aattatttca
241 cgtttgccat taccggttct ccatagggtg atgttcatta gcagtggtga taggttaatt
301 ttcaccatct cttatgcggt tgaatagtca cctctgaacc actttttcct ccagtaactc
361 ctctttcttc ggaccttctg cagccaacct gaaagaataa caaggaggtg gctggaaact
421 tgttttaagg aaccgcctgt ccttcccccg ctggaaacct tgcacctcgg acgctcctgc
481 tcctgccccc acctgacccc cgccctcgtt gacatccagg cgcgatgatc tctgctgcca
541 gtagagggca cacttacttt actttcgcaa acctgaacgc gggtgctgcc cagagagggg
601 gcggagggaa agacgctttg cagcaaaatc cagcatagcg attggttgct ccccgcgttt
661 gcggcaaagg cctggaggca ggagtaattt gcaatcctta aagctgaatt gtgcagtgca
721 tcggatttgg aagctactat attcacttaa cacttgaacg ctgagctgca aactcaacgg
781 gtaataaccc atcttgaaca gcgtacatgc tatacacgca cccctttccc ccgaattgtt
841 ttctcttttg gaggtggtgg agggagagaa aagtttactt aaaatgcctt tgggtgaggg
901 accaaggatg agaagaatgt tttttgtttt tcatgccgtg gaataacaca aaataaaaaa
961 tcccgaggga atatacatta tatattaaat atagatcatt tcagggagca aacaaatcat
1021 gtgtggggct gggcaactag ctaagtcgaa gcgtaaataa aatgtgaata cacgtttgcg
1081 ggttacatac agtgcacttt cactagtatt cagaaaaaat tgtgagtcag tgaactagga
1141 aattaatgcc tggaaggcag ccaaatttta attagctcaa gactcccccc cccccaaaaa
1201 aaggcacgga agtaatactc ctctcctctt ctttgatcag aatcgatgca ttttttgtgc
1261 atgaccgcat ttccaataat aaaaggggaa agaggacctg gaaaggaatt aaacgtccgg
1321 tttgtccggg gaggaaagag ttaacggttt ttttcacaag ggtctctgct gactcccccg
1381 gctcggtcca caagctctcc acttgcccct tttaggaagt ccggtcccgc ggttcgggta
1441 ccccctgccc ctcccatatt ctcccgtcta gcacctttga tttctcccaa acccggcagc
1501 ccgagactgt tgcaaaccgg cgccacaggg cgcaaagggg atttgtctct tctgaaacct
1561 ggctgagaaa ttgggaactc cgtgtgggag gcgtgggggt gggacggtgg ggtacagact
1621 ggcagagagc aggcaacctc cctctcgccc tagcccagct ctggaacagg cagacacatc
1681 tcagggctaa acagacgcct cccgcacggg gccccacgga agcctgagca ggcggggcag
1741 gaggggcggt atctgctgct ttggcagcaa attgggggac tcagtctggg tggaaggtat
1801 ccaatccaga tagctgtgca tacataatgc ataatacatg actcccccca acaaatgcaa
1861 tgggagttta ttcataacgc gctctccaag tatacgtggc aatgcgttgc tgggttattt
1921 taatcattct aggcatcgtt ttcctcctta tgcctctatc attcctccct atctacacta
1981 acatcccacg ctctgaacgc gcgcccatta atacccttct ttcctccact ctccctggga
2041 ctcttgatca aagcgcggcc ctttccccag ccttagcgag gcgccctgca gcctggtacg
2101 cgcgtggcgt ggcggtgggc gcgcagtgcg ttctcggtgt ggagggcagc tgttccgcct
2161 gcgatgattt atactcacag gacaaggatg cggtttgtca aacagtactg ctacggagga
2221 gcagcagaga aagggagagg gtttgagagg gagcaaaaga aaatggtagg cgcgcgtagt
2281 taattcatgc ggctctctta ctctgtttac atcctagagc tagagtgctc ggctgcccgg
2341 ctgagtctcc tccccacctt ccccaccctc cccaccctcc ccataagcgc ccctcccggg
2401 ttcccaaagc agagggcgtg ggggaaaaga aaaaagatcc tctctcgcta atctccgccc
2461 accggccctt tataatgcga gggtctggac ggctgaggac ccccgagctg tgctgctcgc
2521 ggccgccacc gccgggcccc ggccgtccct ggctcccctc ctgcctcgag aagggcaggg
2581 cttctcagag gcttggcggg aaaaagaacg gagggaggga tcgcgctgag tataaaagcc
2641 ggttttcggg gctttatcta actcgctgta gtaattccag cgagag
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