引物怎么和CDNA序列不匹配啊?请大家帮我看看!谢谢
丁香园论坛
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大家好!我打算做半定量RT-PCR,把在文献上查到的引物(用来做实时定量)与GENEBANK里查到的cdna序列用PP5比较了一下,发现怎么好像不匹配?是不是我配的有问题?请大家帮我看看,谢谢!扩增产物有多长?
CDNA:
1 tggcgttgta tagttgggta cagcgaggct ggcgctgcgg tcagacgtgg gcgcctctct
61 tgggtggcgg ctaccgggag ctctctgcga cccaggcccg gcagctgctt ggacgcaggt
121 ttaacctact tcttcagcaa aaatgtggat ttagaaaagc accgaggaag gtggagcctc
181 gaaggtcaga cacagggtca agtggagaag catacaagag aagtgccttg atcccacccc
241 tggaggaaac agtcttttac ccttccccct accctttcac aggctgtgca tttggttcag
301 ctgctatttg gcagtatgaa tcactcaagt ccagggttca gagttatttt gatggcataa
361 aagctgactg gttggatagc atacggccac aaaaggaagg aaacctcaga aaggagatta
421 acaagtggtg gaatagccta agtgatggcc agcggactgt gacaggtatc atagctgcga
481 atgccctcgt attctgctta tggagagtgc cgtccctgca tcgaaccatg atcagatact
541 ttacatccaa cccagcatca aaggttcttt gttccccaat gctgctttcc acgttcagtc
601 atttctcctt attccacatg gcagctaaca tgtatgtttt gtggagcttc tcaactagca
661 tcgtgaacat tctgggtcag gagcagtttg tggcagtgta cctgtctgca ggtgttattt
721 cgaattttgt cagttatgtg tgcaaagttg ccacaggaag atacggacct tcacttggtg
781 cgtcaggcgc aatcatgact gtgcttgcag ctgtctgcac caagatcccg gaggggagac
841 tcgccatcat cttcctccct gttttcacct tcactgcagg caatgcctta aaagccatca
901 ttgccatgga tacagctggg atgatcctgg gatggaagtt ttttgatcat gcagcacatc
961 ttgggggagc cctctttgga atatggtata tcacgtatgg acatgaactc atttggaaga
1021 acagggagcc tctagtgaaa atctggcacg aaataaggac taatggcccc aagaaaggag
1081 gtggctctaa gtgaggcaaa gctggcctgt ctggtggtgc atctgatcct tgttgctcgg
1141 aagtctcagc atcggtcacc ccggtgattt aaatgtcctc agtacgtctc cttcaaaaat
1201 tatggcagag ttgtgaaata aatgtttata tctctagttt gtaaaaaaaa aaaaaaaaaa
1261 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aa
引物:
forward, 5 -ccc acc tct gga aga aac tgt ct-3 ;
reverse 5 - cgg aca ctt ggg ttg tca ctt ct-3
在线急等,谢谢各位老师!
CDNA:
1 tggcgttgta tagttgggta cagcgaggct ggcgctgcgg tcagacgtgg gcgcctctct
61 tgggtggcgg ctaccgggag ctctctgcga cccaggcccg gcagctgctt ggacgcaggt
121 ttaacctact tcttcagcaa aaatgtggat ttagaaaagc accgaggaag gtggagcctc
181 gaaggtcaga cacagggtca agtggagaag catacaagag aagtgccttg atcccacccc
241 tggaggaaac agtcttttac ccttccccct accctttcac aggctgtgca tttggttcag
301 ctgctatttg gcagtatgaa tcactcaagt ccagggttca gagttatttt gatggcataa
361 aagctgactg gttggatagc atacggccac aaaaggaagg aaacctcaga aaggagatta
421 acaagtggtg gaatagccta agtgatggcc agcggactgt gacaggtatc atagctgcga
481 atgccctcgt attctgctta tggagagtgc cgtccctgca tcgaaccatg atcagatact
541 ttacatccaa cccagcatca aaggttcttt gttccccaat gctgctttcc acgttcagtc
601 atttctcctt attccacatg gcagctaaca tgtatgtttt gtggagcttc tcaactagca
661 tcgtgaacat tctgggtcag gagcagtttg tggcagtgta cctgtctgca ggtgttattt
721 cgaattttgt cagttatgtg tgcaaagttg ccacaggaag atacggacct tcacttggtg
781 cgtcaggcgc aatcatgact gtgcttgcag ctgtctgcac caagatcccg gaggggagac
841 tcgccatcat cttcctccct gttttcacct tcactgcagg caatgcctta aaagccatca
901 ttgccatgga tacagctggg atgatcctgg gatggaagtt ttttgatcat gcagcacatc
961 ttgggggagc cctctttgga atatggtata tcacgtatgg acatgaactc atttggaaga
1021 acagggagcc tctagtgaaa atctggcacg aaataaggac taatggcccc aagaaaggag
1081 gtggctctaa gtgaggcaaa gctggcctgt ctggtggtgc atctgatcct tgttgctcgg
1141 aagtctcagc atcggtcacc ccggtgattt aaatgtcctc agtacgtctc cttcaaaaat
1201 tatggcagag ttgtgaaata aatgtttata tctctagttt gtaaaaaaaa aaaaaaaaaa
1261 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aa
引物:
forward, 5 -ccc acc tct gga aga aac tgt ct-3 ;
reverse 5 - cgg aca ctt ggg ttg tca ctt ct-3
在线急等,谢谢各位老师!