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请各位同道用blast帮我分析一下我的引物是否合适。急!!!先谢谢大家了

丁香园论坛

629
AIB1-U primer (5'-ATA CTT GCT GGA TGG TGG ACT-3'; sense; positions 110–130; GenBank accession no. AF012108)
AIB1-L primer (5'-TCC TTG CTC TTT TAT TTG ACG-3'; antisense; positions 458–438; GenBank accession no. AF012108);
如上为我的目的基因上下游引物,请各位同道帮忙用软件及blast分析一下,本人非常焦急,因试验结果很不好,正在考虑是否更换引物!先谢谢大家了!
如下为我的我的目的基因的基因组序列
1 cggcggcggc tgcggcttag tcggtggcgg ccggcggcgg ctgcgggctg agcggcgagt
61 ttccgattta aagctgagct gcgaggaaaa tggcggcggg aggatcaaaa tacttgctgg
121 atggtggact cagagaccaa taaaaataaa ctgcttgaac atcctttgac tggttagcca
181 gttgctgatg tatattcaag atgagtggat taggagaaaa cttggatcca ctggccagtg
241 attcacgaaa acgcaaattg ccatgtgata ctccaggaca aggtcttacc tgcagtggtg
301 aaaaacggag acgggagcag gaaagtaaat atattgaaga attggctgag ctgatatctg
361 ccaatcttag tgatattgac aatttcaatg tcaaaccaga taaatgtgcg attttaaagg
421 aaacagtaag acagatacgt caaataaaag agcaaggaaa aactatttcc aatgatgatg
481 atgttcaaaa agccgatgta tcttctacag ggcagggagt tattgataaa gactccttag
541 gaccgctttt acttcaggca ttggatggtt tcctatttgt ggtgaatcga gacggaaaca
601 ttgtatttgt atcagaaaat gtcacacaat acctgcaata taagcaagag gacctggtta
661 acacaagtgt ttacaatatc ttacatgaag aagacagaaa ggattttctt aagaatttac
721 caaaatctac agttaatgga gtttcctgga caaatgagac ccaaagacaa aaaagccata
781 catttaattg ccgtatgttg atgaaaacac cacatgatat tctggaagac ataaacgcca
841 gtcctgaaat gcgccagaga tatgaaacaa tgcagtgctt tgccctgtct cagccacgag
901 ctatgatgga ggaaggggaa gatttgcaat cttgtatgat ctgtgtggca cgccgcatta
961 ctacaggaga aagaacattt ccatcaaacc ctgagagctt tattaccaga catgatcttt
1021 caggaaaggt tgtcaatata gatacaaatt cactgagatc ctccatgagg cctggctttg
1081 aagatataat ccgaaggtgt attcagagat tttttagtct aaatgatggg cagtcatggt
1141 cccagaaacg tcactatcaa gaagcttatc ttaatggcca tgcagaaacc ccagtatatc
1201 gattctcgtt ggctgatgga actatagtga ctgcacagac aaaaagcaaa ctcttccgaa
1261 atcctgtaac aaatgatcga catggctttg tctcaaccca cttccttcag agagaacaga
1321 atggatatag accaaaccca aatcctgttg gacaagggat tagaccacct atggctggat
1381 gcaacagttc ggtaggcggc atgagtatgt cgccaaacca aggcttacag atgccgagca
1441 gcagggccta tggcttggca gaccctagca ccacagggca gatgagtgga gctaggtatg
1501 ggggttccag taacatagct tcattgaccc ctgggccagg catgcaatca ccatcttcct
1561 accagaacaa caactatggg ctcaacatga gtagcccccc acatgggagt cctggtcttg
1621 ccccaaacca gcagaatatc atgatttctc ctcgtaatcg tgggagtcca aagatagcct
1681 cacatcagtt ttctcctgtt gcaggtgtgc actctcccat ggcatcttct ggcaatactg
1741 ggaaccacag cttttccagc agctctctca gtgccctgca agccatcagt gaaggtgtgg
1801 ggacttccct tttatctact ctgtcatcac caggccccaa attggataac tctcccaata
1861 tgaatattac ccaaccaagt aaagtaagca atcaggattc caagagtcct ctgggctttt
1921 attgcgacca aaatccagtg gagagttcaa tgtgtcagtc aaatagcaga gatcacctca
1981 gtgacaaaga aagtaaggag agcagtgttg agggggcaga gaatcaaagg ggtcctttgg
2041 aaagcaaagg tcataaaaaa ttactgcagt tacttacctg ttcttctgat gaccggggtc
2101 attcctcctt gaccaactcc cccctagatt caagttgtaa agaatcttct gttagtgtca
2161 ccagcccctc tggagtctcc tcctctacat ctggaggagt atcctctaca tccaatatgc
2221 atgggtcact gttacaagag aagcaccgga ttttgcacaa gttgctgcag aatgggaatt
2281 caccagctga ggtagccaag attactgcag aagccactgg gaaagacacc agcagtataa
2341 cttcttgtgg ggacggaaat gttgtcaagc aggagcagct aagtcctaag aagaaggaga
2401 ataatgcact tcttagatac ctgctggaca gggatgatcc tagtgatgca ctctctaaag
2461 aactacagcc ccaagtggaa ggagtggata ataaaatgag tcagtgcacc agctccacca
2521 ttcctagctc aagtcaagag aaagacccta aaattaagac agagacaagt gaagagggat
2581 ctggagactt ggataatcta gatgctattc ttggtgatct gactagttct gacttttaca
2641 ataattccat atcctcaaat ggtagtcatc tggggactaa gcaacaggtg tttcaaggaa
2701 ctaattctct gggtttgaaa agttcacagt ctgtgcagtc tattcgtcct ccatataacc
2761 gagcagtgtc tctggatagc cctgtttctg ttggctcaag tcctccagta aaaaatatca
2821 gtgctttccc catgttacca aagcaaccca tgttgggtgg gaatccaaga atgatggata
2881 gtcaggaaaa ttatggctca agtatgggtg ggccaaaccg aaatgtgact gtgactcaga
2941 ctccttcctc aggagactgg ggcttaccaa actcaaaggc cggcagaatg gaacctatga
3001 attcaaactc catgggaaga ccaggaggag attataatac ttctttaccc agacctgcac
3061 tgggtggctc tattcccaca ttgcctcttc ggtctaatag cataccaggt gcgagaccag
3121 tattgcaaca gcagcagcag atgcttcaaa tgaggcctgg tgaaatcccc atgggaatgg
3181 gggctaatcc ctatggccaa gcagcagcat ctaaccaact gggttcctgg cccgatggca
3241 tgttgtccat ggaacaagtt tctcatggca ctcaaaatag gcctcttctt aggaattccc
3301 tggatgatct tgttgggcca ccttccaacc tggaaggcca gagtgacgaa agagcattat
3361 tggaccagct gcacactctt ctcagcaaca cagatgccac aggcctggaa gaaattgaca
3421 gagctttggg cattcctgaa cttgtcaatc agggacaggc attagagccc aaacaggatg
3481 ctttccaagg ccaagaagca gcagtaatga tggatcagaa ggcaggatta tatggacaga
3541 catacccagc acaggggcct ccaatgcaag gaggctttca tcttcaggga caatcaccat
3601 cttttaactc tatgatgaat cagatgaacc agcaaggcaa ttttcctctc caaggaatgc
3661 acccacgagc caacatcatg agaccccgga caaacacccc caagcaactt agaatgcagc
3721 ttcagcagag gctgcagggc cagcagtttt tgaatcagag ccgacaggca cttgaattga
3781 aaatggaaaa ccctactgct ggtggtgctg cggtgatgag gcctatgatg cagccccagc
3841 agggttttct taatgctcaa atggtcgccc aacgcagcag agagctgcta agtcatcact
3901 tccgacaaca gagggtggct atgatgatgc agcagcagca gcagcagcaa cagcagcagc
3961 agcagcagca gcagcagcaa cagcaacagc aacagcaaca gcagcaacag cagcaaaccc
4021 aggccttcag cccacctcct aatgtgactg cttcccccag catggatggg cttttggcag
4081 gacccacaat gccacaagct cctccgcaac agtttccata tcaaccaaat tatggaatgg
4141 gacaacaacc agatccagcc tttggtcgag tgtctagtcc tcccaatgca atgatgtcgt
4201 caagaatggg tccctcccag aatcccatga tgcaacaccc gcaggctgca tccatctatc
4261 agtcctcaga aatgaagggc tggccatcag gaaatttggc caggaacagc tccttttccc
4321 agcagcagtt tgcccaccag gggaatcctg cagtgtatag tatggtgcac atgaatggca
4381 gcagtggtca catgggacag atgaacatga accccatgcc catgtctggc atgcctatgg
4441 gtcctgatca gaaatactgc tgacatctct gcaccaggac ctcttaagga aaccactgta
4501 caaatgacac tgcactagga ttattgggaa ggaatcattg ttccaggcat ccatcttgga
4561 agaaaggacc agctttgagc tccatcaagg gtattttaag tgatgtcatt tgagcaggac
4621 tggattttaa gccgaagggc aatatctacg tgtttttccc ccctccttct gctgtgtatc
4681 atggtgttca aaacagaaat gttttttggc attccacctc ctagggatat aattctggag
4741 acatggagtg ttactgatca taaaactttt gtgtcacttt tttctgcctt gctagccaaa
4801 atctcttaaa tacacgtagg tgggccagag aacattggaa gaatcaagag agattagaat
4861 atctggtttc tctagttgca gtattggaca aagagcatag tcccagcctt caggtgtagt
4921 agttctgtgt tgaccctttg tccagtggaa ttggtgattc tgaattgtcc tttactaatg
4981 gtgttgagtt gctctgtccc tattatttgc cctaggcttt ctcctaatga aggttttcat
5041 ttgccattca tgtcctgtaa tacttcacct ccaggaactg tcatggatgt ccaaatggct
5101 ttgcagaaag gaaatgagat gacagtattt aatcgcagca gtagcaaact tttcacatgc
5161 taatgtgcag ctgagtgcac tttatttaaa aagaatggat aaatgcaata ttcttgaggt
5221 cttgagggaa tagtgaaaca cattcctggt ttttgcctac acttacgtgt tagacaagaa
5281 ctatgatttt ttttttaaag tactggtgtc accctttgcc tatatggtag agcaataatg
5341 ctttttaaaa ataaacttct gaaaacccaa ggccaggtac tgcattctga atcagaatct
5401 cgcagtgttt ctgtgaatag atttttttgt aaatatgacc tttaagatat tgtattatgt
5461 aaaatatgta tatacctttt tttgtaggtc acaacaactc atttttacag agtttgtgaa
5521 gctaaatatt taacattgtt gatttcagta agctgtgtgg tgaggctacc agtggaagag
5581 acatcccttg acttttgtgg cctgggggag gggtagtgct ccacagcttt tccttcccca
5641 ccccccagcc ttagatgcct cgctcttttc aatctcttaa tctaaatgct ttttaaagag
5701 attatttgtt tagatgtagg cattttaatt ttttaaaaat tcctctacca gaactaagca
5761 ctttgttaat ttggggggaa agaatagata tggggaaata aacttaaaaa aaaatcagga
5821 atttaaaaaa acgagcaatt tgaagagaat cttttggatt ttaagcagtc cgaaataata
5881 gcaattcatg ggctgtgtgt gtgtgtgtat gtgtgtgtgt gtgtgtgtat gtttaattat
5941 gttacctttt catccccttt aggagcgttt tcagattttg gttgctaaga cctgaatccc
6001 atattgagat ctcgagtaga atccttggtg tggtttctgg tgtctgctca gctgtcccct
6061 cattctacta atgtgatgct ttcattatgt ccctgtggat tagaatagtg tcagttattt
6121 cttaagtaac tcagtaccca gaacagccag ttttactgtg attcagagcc acagtctaac
6181 tgagcacctt ttaaacccct ccctcttctg ccccctacca cttttctgct gttgcctctc
6241 tttgacacct gttttagtca gttgggagga agggaaaaat caagtttaat tccctttatc
6301 tgggttaatt catttggttc aaatagttga cggaattggg tttctgaatg tctgtgaatt
6361 tcagaggtct ctgctagcct tggtatcatt ttctagcaat aactgagagc cagttaattt
6421 taagaatttc acacatttag ccaatctttc tagatgtctc tgaaggtaag atcatttaat
6481 atctttgata tgcttacgag taagtgaatc ctgattattt ccagacccac caccagagtg
6541 gatcttattt tcaaagcagt atagacaatt atgagtttgc cctctttccc ctaccaagtt
6601 caaaatatat ctaagaaaga ttgtaaatcc gaaaacttcc attgtagtgg cctgtgcttt
6661 tcagatagta tactctcctg tttggagaca gaggaagaac caggtcagtc tgtctctttt
6721 tcagctcaat tgtatctgac ccttctttaa gttatgtgtg tggggagaaa tagaatggtg
6781 ctcttatctt tcttgacttt aaaaaaatta ttaaaaacaa aaaaaaaaaa aaaaa
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