哪一个引物好啊？
丁香园论坛
1223
准备用RT-PCR扩Sema3b（NM_004636） .
1 gggcgggggc tgcttcttaa aggagccgtc agagggtccc cagcggcccc agggtgggaa
61 ggggccagag tggagagatg ctgctgcgga agtcctcggt ggagtgtgag aaggcagcca
121 gtgttggcct ggaagacctc tagaacctga gaagaggcag cgatcctggg gcagagtcca
181 gggcagctca aggctcctcc acacacacac ccgctgaacc ctgagcaccc tgagctgctg
241 agatggggcg ggccggggct gccgccgtga tcccgggcct ggccctgctc tgggcagtgg
301 ggctggggag tgccgccccc agccccccac gccttcggct ctccttccaa gagctccagg
361 cctggcatgg tctccagact ttcagcctgg agcgaacctg ctgctaccag gccttgctgg
421 tggatgagga gcgtggacgc ctgtttgtgg gtgccgagaa ccatgtggcc tccctcaacc
481 tggacaacat cagcaagcgg gccaagaagc tggcctggcc ggcccctgtg gaatggcgag
541 aggagtgcaa ctgggcaggg aaggacattg gtactgagtg catgaacttc gtgaagttgc
601 tgcatgccta caaccgcacc catttgctgg cctgtggcac gggagccttc cacccaacct
661 gtgcctttgt ggaagtgggc caccgggcag aggagcccgt cctccggctg gacccaggaa
721 ggatagagga tggcaagggg aagagtcctt atgaccccag gcatcgggct gcctccgtgc
781 tggtggggga ggagctatac tcaggggtgg cagcagacct catgggacga gactttacca
841 tctttcgcag cctagggcaa cgtccaagtc tccgaacaga gccacacgac tcccgctggc
901 tcaatgagcc caagtttgtc aaggtatttt ggatcccgga gagcgagaac ccagacgacg
961 acaaaatcta cttcttcttt cgtgagacgg cggtagaggc ggcgccggca ctgggacgcc
1021 tgtccgtgtc ccgcgttggc cagatctgcc ggaacgacgt gggcggccag cgcagcctgg
1081 tcaacaagtg gacgacgttc ctgaaggcgc ggctggtgtg ctcggtgccc ggcgtcgagg
1141 gcgacaccca cttcgatcag ctccaggatg tgtttctgtt gtcctcgcgg gaccaccgga
1201 ccccgctgct ctatgccgtc ttctccacgt ccagcagcat cttccagggc tctgcggtgt
1261 gcgtgtacag catgaacgac gtgcgccggg ccttcttggg accctttgca cacaaggagg
1321 ggcccatgca ccagtgggtg tcataccagg gtcgcgtccc ctacccgcgg ccaggcatgt
1381 gccccagcaa gacctttggc accttcagtt ccaccaagga cttcccagac gatgtcatcc
1441 agtttgcgcg gaaccacccc ctcatgtaca actctgtcct gcccactggg gggcgccctc
1501 ttttcctaca agttggagcc aattacacct tcactcaaat tgccgcggac cgggttgcag
1561 ccgctgacgg acactatgac gtcctcttca ttggcacaga cgttggcacg gtgctgaagg
1621 tgatctcggt ccccaagggc agtaggccca gcgcagaggg gctgctcctg gaggagctgc
1681 acgtgtttga ggactcggcc gctgtcacca gcatgcaaat ttcttccaag aggcaccagc
1741 tgtacgtagc ctcgcggagc gcggtggccc agatcgcgtt gcaccgctgc gctgcccacg
1801 gccgcgtctg caccgaatgc tgtctggcgc gtgaccccta ctgcgcctgg gacggggtcg
1861 cgtgcacgcg cttccagccc agtgccaaga ggcggttccg gcggcaagac gtaaggaatg
1921 gcgaccccag cacgttgtgc tccggagact cgtctcgtcc cgcgctgctg gaacacaagg
1981 tgttcggcgt ggagggcagc agcgcctttc tggagtgtga gccccgctcg ctgcaggcgc
2041 gcgtggagtg gactttccag cgcgcagggg tgacagccca cacccaggtg ctggcagagg
2101 agcgcaccga gcgcaccgcc cggggactac tgctgcgcag gctgcggcgc cgggactcgg
2161 gcgtgtactt gtgcgccgcc gtcgagcagg gctttacgca accgctgcgt cgcctgtcgc
2221 tgcacgtgtt gagtgctacg caggccgaac gactggcgcg ggccgaggag gctgcgcccg
2281 ccgcgccgcc gggccccaaa ctctggtacc gggactttct gcagctggtg gagccgggcg
2341 gaggtggcag cgcgaactcc ctgcgcatgt gccgcccgca gcctgcgctg cagtcactgc
2401 ccctggagtc gcggagaaag ggccgtaacc ggaggaccca cgcccctgag cctcgcgctg
2461 agcgggggcc gcgcagcgca acgcactggt gaccagactg tccccacgcc gggaaccaag
2521 caggagacga caggcgagag aggagccaga cagaccctga aaagaaggac gggttggggc
2581 cgggcacatt gggggtcacc ggccgatgga gacaccaacc gacaggccct ggctgagggc
2641 agctgcgcgg gcttatttat taacaggata acccttgaat gtagcagccc cgggagggcg
2701 gcacaggtcg ggcgcaggat tcagccggag ggaagggacg gggaagccga gctccagagc
2761 aacgaccagg gccgaggagg tgcctggagt gcccaccctg ggagacagac cccacctcct
2821 tgggtagtga gcagtgagca gaaagctgtg aacaggctgg gctgctggag gtggggcgag
2881 gcaggccgac tgtactaaag taacgcaata aacgcattat cagccaaaaa aaaaaaaaaa
2941 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaa
用primer5.0设计了两队引物，不知道哪一个好？评分都是100
引物1：5－GATGGCAAGGGGAAGAGT－3
3－CGAGATACGGCAGAAGAGG－5
1 gggcgggggc tgcttcttaa aggagccgtc agagggtccc cagcggcccc agggtgggaa
61 ggggccagag tggagagatg ctgctgcgga agtcctcggt ggagtgtgag aaggcagcca
121 gtgttggcct ggaagacctc tagaacctga gaagaggcag cgatcctggg gcagagtcca
181 gggcagctca aggctcctcc acacacacac ccgctgaacc ctgagcaccc tgagctgctg
241 agatggggcg ggccggggct gccgccgtga tcccgggcct ggccctgctc tgggcagtgg
301 ggctggggag tgccgccccc agccccccac gccttcggct ctccttccaa gagctccagg
361 cctggcatgg tctccagact ttcagcctgg agcgaacctg ctgctaccag gccttgctgg
421 tggatgagga gcgtggacgc ctgtttgtgg gtgccgagaa ccatgtggcc tccctcaacc
481 tggacaacat cagcaagcgg gccaagaagc tggcctggcc ggcccctgtg gaatggcgag
541 aggagtgcaa ctgggcaggg aaggacattg gtactgagtg catgaacttc gtgaagttgc
601 tgcatgccta caaccgcacc catttgctgg cctgtggcac gggagccttc cacccaacct
661 gtgcctttgt ggaagtgggc caccgggcag aggagcccgt cctccggctg gacccaggaa
721 ggatagagga tggcaagggg aagagtcctt atgaccccag gcatcgggct gcctccgtgc
781 tggtggggga ggagctatac tcaggggtgg cagcagacct catgggacga gactttacca
841 tctttcgcag cctagggcaa cgtccaagtc tccgaacaga gccacacgac tcccgctggc
901 tcaatgagcc caagtttgtc aaggtatttt ggatcccgga gagcgagaac ccagacgacg
961 acaaaatcta cttcttcttt cgtgagacgg cggtagaggc ggcgccggca ctgggacgcc
1021 tgtccgtgtc ccgcgttggc cagatctgcc ggaacgacgt gggcggccag cgcagcctgg
1081 tcaacaagtg gacgacgttc ctgaaggcgc ggctggtgtg ctcggtgccc ggcgtcgagg
1141 gcgacaccca cttcgatcag ctccaggatg tgtttctgtt gtcctcgcgg gaccaccgga
1201 ccccgctgct ctatgccgtc ttctccacgt ccagcagcat cttccagggc tctgcggtgt
1261 gcgtgtacag catgaacgac gtgcgccggg ccttcttggg accctttgca cacaaggagg
1321 ggcccatgca ccagtgggtg tcataccagg gtcgcgtccc ctacccgcgg ccaggcatgt
1381 gccccagcaa gacctttggc accttcagtt ccaccaagga cttcccagac gatgtcatcc
1441 agtttgcgcg gaaccacccc ctcatgtaca actctgtcct gcccactggg gggcgccctc
1501 ttttcctaca agttggagcc aattacacct tcactcaaat tgccgcggac cgggttgcag
1561 ccgctgacgg acactatgac gtcctcttca ttggcacaga cgttggcacg gtgctgaagg
1621 tgatctcggt ccccaagggc agtaggccca gcgcagaggg gctgctcctg gaggagctgc
1681 acgtgtttga ggactcggcc gctgtcacca gcatgcaaat ttcttccaag aggcaccagc
1741 tgtacgtagc ctcgcggagc gcggtggccc agatcgcgtt gcaccgctgc gctgcccacg
1801 gccgcgtctg caccgaatgc tgtctggcgc gtgaccccta ctgcgcctgg gacggggtcg
1861 cgtgcacgcg cttccagccc agtgccaaga ggcggttccg gcggcaagac gtaaggaatg
1921 gcgaccccag cacgttgtgc tccggagact cgtctcgtcc cgcgctgctg gaacacaagg
1981 tgttcggcgt ggagggcagc agcgcctttc tggagtgtga gccccgctcg ctgcaggcgc
2041 gcgtggagtg gactttccag cgcgcagggg tgacagccca cacccaggtg ctggcagagg
2101 agcgcaccga gcgcaccgcc cggggactac tgctgcgcag gctgcggcgc cgggactcgg
2161 gcgtgtactt gtgcgccgcc gtcgagcagg gctttacgca accgctgcgt cgcctgtcgc
2221 tgcacgtgtt gagtgctacg caggccgaac gactggcgcg ggccgaggag gctgcgcccg
2281 ccgcgccgcc gggccccaaa ctctggtacc gggactttct gcagctggtg gagccgggcg
2341 gaggtggcag cgcgaactcc ctgcgcatgt gccgcccgca gcctgcgctg cagtcactgc
2401 ccctggagtc gcggagaaag ggccgtaacc ggaggaccca cgcccctgag cctcgcgctg
2461 agcgggggcc gcgcagcgca acgcactggt gaccagactg tccccacgcc gggaaccaag
2521 caggagacga caggcgagag aggagccaga cagaccctga aaagaaggac gggttggggc
2581 cgggcacatt gggggtcacc ggccgatgga gacaccaacc gacaggccct ggctgagggc
2641 agctgcgcgg gcttatttat taacaggata acccttgaat gtagcagccc cgggagggcg
2701 gcacaggtcg ggcgcaggat tcagccggag ggaagggacg gggaagccga gctccagagc
2761 aacgaccagg gccgaggagg tgcctggagt gcccaccctg ggagacagac cccacctcct
2821 tgggtagtga gcagtgagca gaaagctgtg aacaggctgg gctgctggag gtggggcgag
2881 gcaggccgac tgtactaaag taacgcaata aacgcattat cagccaaaaa aaaaaaaaaa
2941 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaa
用primer5.0设计了两队引物，不知道哪一个好？评分都是100
引物1：5－GATGGCAAGGGGAAGAGT－3
3－CGAGATACGGCAGAAGAGG－5
