RT-PCR求教！！！
丁香园论坛
797
请大侠帮忙看一下用下列两引物进行PCR扩增有没有可行性
上游引物：5'-GCCAGACTGGGAAGAAATCTG-3’
下游引物：5'-TGTGTTGGAAAATCCAAGTCA-3’
基因序列：
1 aggcagtgga gccccggcgg cggcggcggc ggcgcgcggg ggcgacgcgc gggaacaacg
61 cgagtcggcg cgcgggacga agaataatca tgggccagac tgggaagaaa tctgagaagg
121 gaccagtttg ttggcggaag cgtgtaaaat cagagtacat gcgactgaga cagctcaaga
181 ggttcagacg agctgatgaa gtaaagagta tgtttagttc caatcgtcag aaaattttgg
241 aaagaacgga aatcttaaac caagaatgga aacagcgaag gatacagcct gtgcacatcc
301 tgacttctgt gagctcattg cgcgggacta gggagtgttc ggtgaccagt gacttggatt
361 ttccaacaca agtcatccca ttaaagactc tgaatgcagt tgcttcagta cccataatgt
421 attcttggtc tcccctacag cagaatttta tggtggaaga tgaaactgtt ttacataaca
481 ttccttatat gggagatgaa gttttagatc aggatggtac tttcattgaa gaactaataa
541 aaaattatga tgggaaagta cacggggata gagaatgtgg gtttataaat gatgaaattt
601 ttgtggagtt ggtgaatgcc cttggtcaat ataatgatga tgacgatgat gatgatggag
661 acgatcctga agaaagagaa gaaaagcaga aagatctgga ggatcaccga gatgataaag
721 aaagccgccc acctcggaaa tttccttctg ataaaatttt tgaagccatt tcctcaatgt
781 ttccagataa gggcacagca gaagaactaa aggaaaaata taaagaactc accgaacagc
841 agctcccagg cgcacttcct cctgaatgta cccccaacat agatggacca aatgctaaat
901 ctgttcagag agagcaaagc ttacactcct ttcatacgct tttctgtagg cgatgtttta
961 aatatgactg cttcctacat ccttttcatg caacacccaa cacttataag cggaagaaca
1021 cagaaacagc tctagacaac aaaccttgtg gaccacagtg ttaccagcat ttggagggag
1081 caaaggagtt tgctgctgct ctcaccgctg agcggataaa gaccccacca aaacgtccag
1141 gaggccgcag aagaggacgg cttcccaata acagtagcag gcccagcacc cccaccatta
1201 atgtgctgga atcaaaggat acagacagtg atagggaagc agggactgaa acggggggag
1261 agaacaatga taaagaagaa gaagagaaga aagatgaaac ttcgagctcc tctgaagcaa
1321 attctcggtg tcaaacacca ataaagatga agccaaatat tgaacctcct gagaatgtgg
1381 agtggagtgg tgctgaagcc tcaatgttta gagtcctcat tggcacttac tatgacaatt
1441 tctgtgccat tgctaggtta attgggacca aaacatgtag acaggtgtat gagtttagag
1501 tcaaagaatc tagcatcata gctccagctc ccgctgagga tgtggatact cctccaagga
1561 aaaagaagag gaaacaccgg ttgtgggctg cacactgcag aaagatacag ctgaaaaagg
1621 acggctcctc taaccatgtt tacaactatc aaccctgtga tcatccacgg cagccttgtg
1681 acagttcgtg cccttgtgtg atagcacaaa atttttgtga aaagttttgt caatgtagtt
1741 cagagtgtca aaaccgcttt ccgggatgcc gctgcaaagc acagtgcaac accaagcagt
1801 gcccgtgcta cctggctgtc cgagagtgtg accctgacct ctgtcttact tgtggagccg
1861 ctgaccattg ggacagtaaa aatgtgtcct gcaagaactg cagtattcag cggggctcca
1921 aaaagcatct attgctggca ccatctgacg tggcaggctg ggggattttt atcaaagatc
1981 ctgtgcagaa aaatgaattc atctcagaat actgtggaga gattatttct caagatgaag
2041 ctgacagaag agggaaagtg tatgataaat acatgtgcag ctttctgttc aacttgaaca
2101 atgattttgt ggtggatgca acccgcaagg gtaacaaaat tcgttttgca aatcattcgg
2161 taaatccaaa ctgctatgca aaagttatga tggttaacgg tgatcacagg ataggtattt
2221 ttgccaagag agccatccag actggcgaag agctgttttt tgattacaga tacagccagg
2281 ctgatgccct gaagtatgtc ggcatcgaaa gagaaatgga aatcccttga catctgctac
2341 ctcctccccc tcctctgaaa cagctgcctt agcttcagga acctcgagta ctgtgggcaa
2401 tttagaaaaa gaacatgcag tttgaaattc tgaatttgca aagtactgta agaataattt
2461 atagtaatga gtttaaaaat caacttttta ttgccttctc accagctgca aagtgttttg
2521 taccagtgaa tttttgcaat aatgcagtat ggtacatttt tcaactttga ataaagaata
2581 cttgaacttg tcaaaaaaaa
上游引物：5'-GCCAGACTGGGAAGAAATCTG-3’
下游引物：5'-TGTGTTGGAAAATCCAAGTCA-3’
基因序列：
1 aggcagtgga gccccggcgg cggcggcggc ggcgcgcggg ggcgacgcgc gggaacaacg
61 cgagtcggcg cgcgggacga agaataatca tgggccagac tgggaagaaa tctgagaagg
121 gaccagtttg ttggcggaag cgtgtaaaat cagagtacat gcgactgaga cagctcaaga
181 ggttcagacg agctgatgaa gtaaagagta tgtttagttc caatcgtcag aaaattttgg
241 aaagaacgga aatcttaaac caagaatgga aacagcgaag gatacagcct gtgcacatcc
301 tgacttctgt gagctcattg cgcgggacta gggagtgttc ggtgaccagt gacttggatt
361 ttccaacaca agtcatccca ttaaagactc tgaatgcagt tgcttcagta cccataatgt
421 attcttggtc tcccctacag cagaatttta tggtggaaga tgaaactgtt ttacataaca
481 ttccttatat gggagatgaa gttttagatc aggatggtac tttcattgaa gaactaataa
541 aaaattatga tgggaaagta cacggggata gagaatgtgg gtttataaat gatgaaattt
601 ttgtggagtt ggtgaatgcc cttggtcaat ataatgatga tgacgatgat gatgatggag
661 acgatcctga agaaagagaa gaaaagcaga aagatctgga ggatcaccga gatgataaag
721 aaagccgccc acctcggaaa tttccttctg ataaaatttt tgaagccatt tcctcaatgt
781 ttccagataa gggcacagca gaagaactaa aggaaaaata taaagaactc accgaacagc
841 agctcccagg cgcacttcct cctgaatgta cccccaacat agatggacca aatgctaaat
901 ctgttcagag agagcaaagc ttacactcct ttcatacgct tttctgtagg cgatgtttta
961 aatatgactg cttcctacat ccttttcatg caacacccaa cacttataag cggaagaaca
1021 cagaaacagc tctagacaac aaaccttgtg gaccacagtg ttaccagcat ttggagggag
1081 caaaggagtt tgctgctgct ctcaccgctg agcggataaa gaccccacca aaacgtccag
1141 gaggccgcag aagaggacgg cttcccaata acagtagcag gcccagcacc cccaccatta
1201 atgtgctgga atcaaaggat acagacagtg atagggaagc agggactgaa acggggggag
1261 agaacaatga taaagaagaa gaagagaaga aagatgaaac ttcgagctcc tctgaagcaa
1321 attctcggtg tcaaacacca ataaagatga agccaaatat tgaacctcct gagaatgtgg
1381 agtggagtgg tgctgaagcc tcaatgttta gagtcctcat tggcacttac tatgacaatt
1441 tctgtgccat tgctaggtta attgggacca aaacatgtag acaggtgtat gagtttagag
1501 tcaaagaatc tagcatcata gctccagctc ccgctgagga tgtggatact cctccaagga
1561 aaaagaagag gaaacaccgg ttgtgggctg cacactgcag aaagatacag ctgaaaaagg
1621 acggctcctc taaccatgtt tacaactatc aaccctgtga tcatccacgg cagccttgtg
1681 acagttcgtg cccttgtgtg atagcacaaa atttttgtga aaagttttgt caatgtagtt
1741 cagagtgtca aaaccgcttt ccgggatgcc gctgcaaagc acagtgcaac accaagcagt
1801 gcccgtgcta cctggctgtc cgagagtgtg accctgacct ctgtcttact tgtggagccg
1861 ctgaccattg ggacagtaaa aatgtgtcct gcaagaactg cagtattcag cggggctcca
1921 aaaagcatct attgctggca ccatctgacg tggcaggctg ggggattttt atcaaagatc
1981 ctgtgcagaa aaatgaattc atctcagaat actgtggaga gattatttct caagatgaag
2041 ctgacagaag agggaaagtg tatgataaat acatgtgcag ctttctgttc aacttgaaca
2101 atgattttgt ggtggatgca acccgcaagg gtaacaaaat tcgttttgca aatcattcgg
2161 taaatccaaa ctgctatgca aaagttatga tggttaacgg tgatcacagg ataggtattt
2221 ttgccaagag agccatccag actggcgaag agctgttttt tgattacaga tacagccagg
2281 ctgatgccct gaagtatgtc ggcatcgaaa gagaaatgga aatcccttga catctgctac
2341 ctcctccccc tcctctgaaa cagctgcctt agcttcagga acctcgagta ctgtgggcaa
2401 tttagaaaaa gaacatgcag tttgaaattc tgaatttgca aagtactgta agaataattt
2461 atagtaatga gtttaaaaat caacttttta ttgccttctc accagctgca aagtgttttg
2521 taccagtgaa tttttgcaat aatgcagtat ggtacatttt tcaactttga ataaagaata
2581 cttgaacttg tcaaaaaaaa
