请帮忙看看引物设计可以吗
丁香园论坛
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我要克隆一全长CDNA，设计引物如下
sense:TCTCGGACCATGTCTCCCGCCCCA（起于104）
antisense:CTA TCA GAC CTT GTC CAG CAG GGA（1841－1861）
cDNA全长为：
1 agttcagtgc ctaccgaaga caaaggcgcc ccgagggagt ggcggtgcga ccccagggcg
61 tgggcccggc cgcggagccc acactgcccg gctgacccgg tggtctcgga ccatgtctcc
121 cgccccaaga ccctcccgtt gtctcctgct ccccctgctc acgctcggca ccgcgctcgc
181 ctccctcggc tcggcccaaa gcagcagctt cagccccgaa gcctggctac agcaatatgg
241 ctacctgcct cccggggacc tacgtaccca cacacagcgc tcaccccagt cactctcagc
301 ggccatcgct gccatgcaga agttttacgg cttgcaagta acaggcaaag ctgatgcaga
361 caccatgaag gccatgaggc gcccccgatg tggtgttcca gacaagtttg gggctgagat
421 caaggccaat gttcgaagga agcgctacgc catccagggt ctcaaatggc aacataatga
481 aatcactttc tgcatccaga attacacccc caaggtgggc gagtatgcca catacgaggc
541 cattcgcaag gcgttccgcg tgtgggagag tgccacacca ctgcgcttcc gcgaggtgcc
601 ctatgcctac atccgtgagg gccatgagaa gcaggccgac atcatgatct tctttgccga
661 gggcttccat ggcgacagca cgcccttcga tggtgagggc ggcttcctgg cccatgccta
721 cttcccaggc cccaacattg gaggagacac ccactttgac tctgccgagc cttggactgt
781 caggaatgag gatctgaatg gaaatgacat cttcctggtg gctgtgcacg agctgggcca
841 tgccctgggg ctcgagcatt ccagtgaccc ctcggccatc atggcaccct tttaccagtg
901 gatggacacg gagaattttg tgctgcccga tgatgaccgc cggggcatcc agcaacttta
961 tgggggtgag tcagggttcc ccaccaagat gccccctcaa cccaggacta cctcccggcc
1021 ttctgttcct gataaaccca aaaaccccac ctatgggccc aacatctgtg acgggaactt
1081 tgacaccgtg gccatgctcc gaggggagat gtttgtcttc aaggagcgct ggttctggcg
1141 ggtgaggaat aaccaagtga tggatggata cccaatgccc attggccagt tctggcgggg
1201 cctgcctgcg tccatcaaca ctgcctacga gaggaaggat ggcaaattcg tcttcttcaa
1261 aggagacaag cattgggtgt ttgatgaggc gtccctggaa cctggctacc ccaagcacat
1321 taaggagctg ggccgagggc tgcctaccga caagattgat gctgctctct tctggatgcc
1381 caatggaaag acctacttct tccgtggaaa caagtactac cgtttcaacg aagagctcag
1441 ggcagtggat agcgagtacc ccaagaacat caaagtctgg gaagggatcc ctgagtctcc
1501 cagagggtca ttcatgggca gcgatgaagt cttcacttac ttctacaagg ggaacaaata
1561 ctggaaattc aacaaccaga agctgaaggt agaaccgggc taccccaagc cagccctgag
1621 ggactggatg ggctgcccat cgggaggccg gccggatgag gggactgagg aggagacgga
1681 ggtgatcatc attgaggtgg acgaggaggg cggcggggcg gtgagcgcgg ctgccgtggt
1741 gctgcccgtg ctgctgctgc tcctggtgct ggcggtgggc cttgcagtct tcttcttcag
1801 acgccatggg acccccaggc gactgctcta ctgccagcgt tccctgctgg acaaggtctg
1861 acgcccaccg ccggcccgcc cactcctacc acaaggactt tgcctctgaa ggccagtggc
1921 agcaggtggt ggtgggtggg ctgttcccat cgtcccgagc cccctccccg cagcctcctt
1981 gcttctctct gtcccctggc tggcctcctt caccctgacc gcctccctcc ctcctgcccc
2041 ggcattgcat cttccctaga taggtcccct gagggctgag tgggagggcg gccctttcca
2101 gcctctgccc ctcaggggaa ccctgtagct ttgtgtctgt ccagccccat ctgaatgtgt
2161 tgggggctct gcacttgaag gcaggaccct cagacctcgc tggtaaaggt caaatggggt
2221 catctgctcc ttttccatcc cctgacatac cttaacctct gaactctgac ctcaggaggc
2281 tctgggcact ccagccctga aagccccagg tgtacccaat tggcagcctc tcactactct
2341 ttctggctaa aaggaatcta atcttgttg
有劳大家了，谢谢
sense:TCTCGGACCATGTCTCCCGCCCCA（起于104）
antisense:CTA TCA GAC CTT GTC CAG CAG GGA（1841－1861）
cDNA全长为：
1 agttcagtgc ctaccgaaga caaaggcgcc ccgagggagt ggcggtgcga ccccagggcg
61 tgggcccggc cgcggagccc acactgcccg gctgacccgg tggtctcgga ccatgtctcc
121 cgccccaaga ccctcccgtt gtctcctgct ccccctgctc acgctcggca ccgcgctcgc
181 ctccctcggc tcggcccaaa gcagcagctt cagccccgaa gcctggctac agcaatatgg
241 ctacctgcct cccggggacc tacgtaccca cacacagcgc tcaccccagt cactctcagc
301 ggccatcgct gccatgcaga agttttacgg cttgcaagta acaggcaaag ctgatgcaga
361 caccatgaag gccatgaggc gcccccgatg tggtgttcca gacaagtttg gggctgagat
421 caaggccaat gttcgaagga agcgctacgc catccagggt ctcaaatggc aacataatga
481 aatcactttc tgcatccaga attacacccc caaggtgggc gagtatgcca catacgaggc
541 cattcgcaag gcgttccgcg tgtgggagag tgccacacca ctgcgcttcc gcgaggtgcc
601 ctatgcctac atccgtgagg gccatgagaa gcaggccgac atcatgatct tctttgccga
661 gggcttccat ggcgacagca cgcccttcga tggtgagggc ggcttcctgg cccatgccta
721 cttcccaggc cccaacattg gaggagacac ccactttgac tctgccgagc cttggactgt
781 caggaatgag gatctgaatg gaaatgacat cttcctggtg gctgtgcacg agctgggcca
841 tgccctgggg ctcgagcatt ccagtgaccc ctcggccatc atggcaccct tttaccagtg
901 gatggacacg gagaattttg tgctgcccga tgatgaccgc cggggcatcc agcaacttta
961 tgggggtgag tcagggttcc ccaccaagat gccccctcaa cccaggacta cctcccggcc
1021 ttctgttcct gataaaccca aaaaccccac ctatgggccc aacatctgtg acgggaactt
1081 tgacaccgtg gccatgctcc gaggggagat gtttgtcttc aaggagcgct ggttctggcg
1141 ggtgaggaat aaccaagtga tggatggata cccaatgccc attggccagt tctggcgggg
1201 cctgcctgcg tccatcaaca ctgcctacga gaggaaggat ggcaaattcg tcttcttcaa
1261 aggagacaag cattgggtgt ttgatgaggc gtccctggaa cctggctacc ccaagcacat
1321 taaggagctg ggccgagggc tgcctaccga caagattgat gctgctctct tctggatgcc
1381 caatggaaag acctacttct tccgtggaaa caagtactac cgtttcaacg aagagctcag
1441 ggcagtggat agcgagtacc ccaagaacat caaagtctgg gaagggatcc ctgagtctcc
1501 cagagggtca ttcatgggca gcgatgaagt cttcacttac ttctacaagg ggaacaaata
1561 ctggaaattc aacaaccaga agctgaaggt agaaccgggc taccccaagc cagccctgag
1621 ggactggatg ggctgcccat cgggaggccg gccggatgag gggactgagg aggagacgga
1681 ggtgatcatc attgaggtgg acgaggaggg cggcggggcg gtgagcgcgg ctgccgtggt
1741 gctgcccgtg ctgctgctgc tcctggtgct ggcggtgggc cttgcagtct tcttcttcag
1801 acgccatggg acccccaggc gactgctcta ctgccagcgt tccctgctgg acaaggtctg
1861 acgcccaccg ccggcccgcc cactcctacc acaaggactt tgcctctgaa ggccagtggc
1921 agcaggtggt ggtgggtggg ctgttcccat cgtcccgagc cccctccccg cagcctcctt
1981 gcttctctct gtcccctggc tggcctcctt caccctgacc gcctccctcc ctcctgcccc
2041 ggcattgcat cttccctaga taggtcccct gagggctgag tgggagggcg gccctttcca
2101 gcctctgccc ctcaggggaa ccctgtagct ttgtgtctgt ccagccccat ctgaatgtgt
2161 tgggggctct gcacttgaag gcaggaccct cagacctcgc tggtaaaggt caaatggggt
2221 catctgctcc ttttccatcc cctgacatac cttaacctct gaactctgac ctcaggaggc
2281 tctgggcact ccagccctga aagccccagg tgtacccaat tggcagcctc tcactactct
2341 ttctggctaa aaggaatcta atcttgttg
有劳大家了，谢谢
