帮忙看一下我设计的引物
丁香园论坛
685
我是新手，对设计引物还不太熟悉，下游CG含量高，没办法下手，麻烦大家帮我看一下
部分DNA序列如下：
1 actgaacttc acagggcaag gggagcccga ctccattcgc tgtgacacac gagcggagct
61 gctgtcaaag ggctgcccag ctgatgacat catggaaccc aagagcctcg ctgagacccg
121 ggacagccag gcgggcagtc ggaagcagct gtccccacag gaagtgacgc tctacctgag
181 accaggtagg cttggctggc taggggtggg ccggccctaa tggcgcgtgt atccatgtgt
241 ccctgcccca ggaggatgtc cccaacgccc catctgcagc tggtgcctag tctcgctttt
301 agcactgagt tctgcgtcct aaaaaaaagg gagaataaac catctggaaa acactcctta
361 tccctctctc agaagatgag gtattttttg gctgagtgaa ggctctgata aggcctgtaa
421 actagaaaca caagtaaacc tttgaatcaa tgttgcccac ctgacttaac cacagaccga
481 ggtataggac acagctctct gttagcactc caagaaacat ccttcaggaa ggtggcttgg
541 tggaacagca gggggttatg gtggatggcc atgggcaggg ctggggtagc cacatggggg
601 agctgatgct ggaaggtggg ctggcagagg ggacttgatg gtcatggctt tgtcgagcac
661 caggcatcta cctgagagct ggggacacag gctatttgta ttgacaagga aagaagtcgt
721 tacaagtgca ggtggtgatt aagatgacga ggagcagaaa ctagggcagc tgggattgag
781 gaagtggggg ggggtgtccc aggatggcca gggagccctt ttggggccct ggcagtttgc
841 ttagcagctg gtggtagaga aggccttacc agagagacca gagagatagt agaatgatta
901 acagtgtgat aacatgacac tctatatctc tgtatccagc acatatgtat ccagatgttt
961 ctatgtcaga acgtgtgctt gcctgaaatg gaatctgaat aggcatcctg catcatatcc
1021 accagcataa gagaatgggg agagtcctga ggttctgagg cctgacaaga tgccataagt
1081 gcccatgaac cccccccacc cccagaccag atagtacacc ctgactatct cccaaatcct
1141 ggcaggtcag gcagttgcgt tcaacgtgac cttccggagg gccaagggct accccatcga
1201 cctgtactac ctgatggacc tctcctactc catggtggat gacctcgtca acgtcaagaa
1261 gctggggggt gacctgctcc gggccctcaa tggcatcacc gagtcgggcc gcattggtga
1321 ggcagctact ccatcttccc ctgaaacccc aagccccggt cccaggcacc tctgcacctc
1381 tgcaccccaa gggcagggcg ccaggcccct ccccacgtgc cgggccttcc tacccctctc
1441 tggtaagagg ctcacggccc ctcactgtcc cccaggtttc gggtccttcg tggacaagac
1501 ggtgctcccc ttcgtcaaca cgcaccccga gaagctgcgg aacccctgcc ccaacaagga
1561 gaaggagtgc cagcccccgt tcgccttcag gcacgtgttg aagctcactg acaactccaa
1621 acagttcgag acagaagtcg
1146到1316是外显子区，两边是内含子区，目的片段是外显子区，1200处为突变位点,要做酶切,想从外显子两边设计引物，扩增片段长度大约在300到400bp，帮忙分析一下我用PRIMER设计的引物,
上游:5'-CCA GCA TAA GAG AAT GGG GAG AG
下游:5'-CCC AAA GTC CCC TTC TAC CT
各位帮我看一下吧
部分DNA序列如下：
1 actgaacttc acagggcaag gggagcccga ctccattcgc tgtgacacac gagcggagct
61 gctgtcaaag ggctgcccag ctgatgacat catggaaccc aagagcctcg ctgagacccg
121 ggacagccag gcgggcagtc ggaagcagct gtccccacag gaagtgacgc tctacctgag
181 accaggtagg cttggctggc taggggtggg ccggccctaa tggcgcgtgt atccatgtgt
241 ccctgcccca ggaggatgtc cccaacgccc catctgcagc tggtgcctag tctcgctttt
301 agcactgagt tctgcgtcct aaaaaaaagg gagaataaac catctggaaa acactcctta
361 tccctctctc agaagatgag gtattttttg gctgagtgaa ggctctgata aggcctgtaa
421 actagaaaca caagtaaacc tttgaatcaa tgttgcccac ctgacttaac cacagaccga
481 ggtataggac acagctctct gttagcactc caagaaacat ccttcaggaa ggtggcttgg
541 tggaacagca gggggttatg gtggatggcc atgggcaggg ctggggtagc cacatggggg
601 agctgatgct ggaaggtggg ctggcagagg ggacttgatg gtcatggctt tgtcgagcac
661 caggcatcta cctgagagct ggggacacag gctatttgta ttgacaagga aagaagtcgt
721 tacaagtgca ggtggtgatt aagatgacga ggagcagaaa ctagggcagc tgggattgag
781 gaagtggggg ggggtgtccc aggatggcca gggagccctt ttggggccct ggcagtttgc
841 ttagcagctg gtggtagaga aggccttacc agagagacca gagagatagt agaatgatta
901 acagtgtgat aacatgacac tctatatctc tgtatccagc acatatgtat ccagatgttt
961 ctatgtcaga acgtgtgctt gcctgaaatg gaatctgaat aggcatcctg catcatatcc
1021 accagcataa gagaatgggg agagtcctga ggttctgagg cctgacaaga tgccataagt
1081 gcccatgaac cccccccacc cccagaccag atagtacacc ctgactatct cccaaatcct
1141 ggcaggtcag gcagttgcgt tcaacgtgac cttccggagg gccaagggct accccatcga
1201 cctgtactac ctgatggacc tctcctactc catggtggat gacctcgtca acgtcaagaa
1261 gctggggggt gacctgctcc gggccctcaa tggcatcacc gagtcgggcc gcattggtga
1321 ggcagctact ccatcttccc ctgaaacccc aagccccggt cccaggcacc tctgcacctc
1381 tgcaccccaa gggcagggcg ccaggcccct ccccacgtgc cgggccttcc tacccctctc
1441 tggtaagagg ctcacggccc ctcactgtcc cccaggtttc gggtccttcg tggacaagac
1501 ggtgctcccc ttcgtcaaca cgcaccccga gaagctgcgg aacccctgcc ccaacaagga
1561 gaaggagtgc cagcccccgt tcgccttcag gcacgtgttg aagctcactg acaactccaa
1621 acagttcgag acagaagtcg
1146到1316是外显子区，两边是内含子区，目的片段是外显子区，1200处为突变位点,要做酶切,想从外显子两边设计引物，扩增片段长度大约在300到400bp，帮忙分析一下我用PRIMER设计的引物,
上游:5'-CCA GCA TAA GAG AAT GGG GAG AG
下游:5'-CCC AAA GTC CCC TTC TAC CT
各位帮我看一下吧
