请帮我设计RT-PCR引物（已有基因原序列）
丁香园论坛
993
请高手帮忙设计一对引物用于RT-PCR,有原基因全序列，我查了国外国内的文献上的引物，经PrimerPrimer5.0验证发现都不太好，自己又从来没有设计过引物序列，故恳请各位有经验的高手帮忙设计一下，多谢！！基因序列如下：
ORIGIN
1 gggggggggg atttagagac ttgctcttgc actaccaaag ccacaaagca gccttgcaga
61 aaagagagct ccatcatgcc tggctcagca ctgctatgct gcctgctctt actgactggc
121 atgaggatca gcaggggcca gtacagccgg gaagacaata actgcaccca cttcccagtc
181 ggccagagcc acatgctcct agagctgcgg actgccttca gccaggtgaa gactttcttt
241 caaacaaagg accagctgga caacatactg ctaaccgact ccttaatgca ggactttaag
301 ggttacttgg gttgccaagc cttatcggaa atgatccagt tttacctggt agaagtgatg
361 ccccaggcag agaagcatgg cccagaaatc aaggagcatt tgaattccct gggtgagaag
421 ctgaagaccc tcaggatgcg gctgaggcgc tgtcatcgat ttctcccctg tgaaaataag
481 agcaaggcag tggagcaggt gaagagtgat tttaataagc tccaagacca aggtgtctac
541 aaggccatga atgaatttga catcttcatc aactgcatag aagcatacat gatgatcaaa
601 atgaaaagct aaaacacctg cagtgtgtat tgagtctgct ggactccagg acctagacag
661 agctctctaa atctgatcca gggatcttag ctaacggaaa caactccttg gaaaacctcg
721 tttgtacctc tctccgaaat atttattacc tctgatacct cagttcccat tctatttatt
781 cactgagctt ctctgtgaac tatttagaaa gaagcccaat attataattt tacagtattt
841 attattttta acctgtgttt aagctgtttc cattggggac actttatagt atttaaaggg
901 agattatatt atatgatggg aggggttctt ccttgggaag caattgaagc ttctattcta
961 aggctggcca cacttgagag ctgcagggcc ctttgctatg gtgtcctttc aattgctctc
1021 atccctgagt tcagagctcc taagagagtt gtgaagaaac tcatgggtct tgggaagaga
1081 aaccagggag atcctttgat gatcattcct gcagcagctc agagggttcc cctactgtca
1141 tcccccagcc gcttcatccc tgaaaactgt ggccagtttg ttatttataa ccacctaaaa
1201 ttagttctaa tagaactcat ttttaactag aagtaatgca attcctctgg gaatggtgta
1261 ttgtttgtct gcctttgtag cagcatctaa ttttgaataa atggatctta ttcg
//
ORIGIN
1 gggggggggg atttagagac ttgctcttgc actaccaaag ccacaaagca gccttgcaga
61 aaagagagct ccatcatgcc tggctcagca ctgctatgct gcctgctctt actgactggc
121 atgaggatca gcaggggcca gtacagccgg gaagacaata actgcaccca cttcccagtc
181 ggccagagcc acatgctcct agagctgcgg actgccttca gccaggtgaa gactttcttt
241 caaacaaagg accagctgga caacatactg ctaaccgact ccttaatgca ggactttaag
301 ggttacttgg gttgccaagc cttatcggaa atgatccagt tttacctggt agaagtgatg
361 ccccaggcag agaagcatgg cccagaaatc aaggagcatt tgaattccct gggtgagaag
421 ctgaagaccc tcaggatgcg gctgaggcgc tgtcatcgat ttctcccctg tgaaaataag
481 agcaaggcag tggagcaggt gaagagtgat tttaataagc tccaagacca aggtgtctac
541 aaggccatga atgaatttga catcttcatc aactgcatag aagcatacat gatgatcaaa
601 atgaaaagct aaaacacctg cagtgtgtat tgagtctgct ggactccagg acctagacag
661 agctctctaa atctgatcca gggatcttag ctaacggaaa caactccttg gaaaacctcg
721 tttgtacctc tctccgaaat atttattacc tctgatacct cagttcccat tctatttatt
781 cactgagctt ctctgtgaac tatttagaaa gaagcccaat attataattt tacagtattt
841 attattttta acctgtgttt aagctgtttc cattggggac actttatagt atttaaaggg
901 agattatatt atatgatggg aggggttctt ccttgggaag caattgaagc ttctattcta
961 aggctggcca cacttgagag ctgcagggcc ctttgctatg gtgtcctttc aattgctctc
1021 atccctgagt tcagagctcc taagagagtt gtgaagaaac tcatgggtct tgggaagaga
1081 aaccagggag atcctttgat gatcattcct gcagcagctc agagggttcc cctactgtca
1141 tcccccagcc gcttcatccc tgaaaactgt ggccagtttg ttatttataa ccacctaaaa
1201 ttagttctaa tagaactcat ttttaactag aagtaatgca attcctctgg gaatggtgta
1261 ttgtttgtct gcctttgtag cagcatctaa ttttgaataa atggatctta ttcg
//
