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[求助] 片段验证不出来

我有一个基因的片段,在不同位置设计了引物,可是怎么都验证不出来它,基本上就没P出条带,求解。下面是基因的序列TGGTGATAAATTAGTGAAGCAAATGTTATTGTTCTTTTGTGTTTCAGTAAAAAAAATAAAAATAGCAACACTATGTATATACACTATTTATATAGACGTAATAGTATGTTAAAGTAGTACTAATCATTCTTTGTTTATTTATATTCCTGTCTGTCTCTTCTTATTGCAAAACAGTGAACTATTAATCTAATAAATTATTTAGGGGGTGAAAATCTATTAAAAGTATGAAAAGTTCTGATCAGAAACTGGGGCCCAATTCTCGCGGCATAGATATTATATCACCTTCGATATATATCGAATGATCTGGCTTTTATCGAAGGAAAACCTTTTTGCCCTATTCTCACCATGTCGCAATAGATATTTATCTACGGATGATCAGTAGACATACATCGACTGGTAAAGAGCCTTCGATATATAAATATCTATGGATCTTGGCATGTCAATCTTGTTATTTCCACTTTTTTTACCTATTATTTATTGTGTTGTCGTGTGTCTTTGCCAATTATTGTTACCTCAAGATGTTGTTAACTGCCTTCATAAATAATTAGTAAATAAAGAAGTAGGTAAACAATGGTGTCGATTCTGGCGTGATTCTCTAAACTTAAAACTAAATTTAACATCAGTCTCTCCTTTTCATTCTGTATAGAGAATGACAAGGATACACTGTTGTCAAATTTAAGTTTAGGTTTAGAGAATCATGCTAGAATCGACACCATTGAAAAATCTAGAGACTAGGAAGTTTTATTTACTGGCTGTTTTTCTATTATCTACGTTTCAGATGTATGAAACTGTAGATCTCTTCTCAAGACTTTACACTATACTAATGATTAATATGATGGTCGGAGGAATGATTCTCTTTAATCTGACTCCATTATACAGCAACTATAAAAACGGTGTTTTCTCTAAGAACCCACCCGAAAACGTAACTTATGCGTATTCTGTCACTTACTCAGTACCAGGATTTAATTTTTATGAGCATTTTACACTGACGACAATTTTGAATTGGATCATGAGCTACGACGTTTCAGTAAATGTCTGTGTAAAAGATCTGTATTTATCATTTCTTGTATTTCAAATAATTGGCCACATACAAATCTTAAAATACAATCTAGAACATTTTCCTAAGCCTAAAAATCAAGCGACTAATCGTTTTGACGCTGAAGAGAATAAGCAGATACGAAAAACGATCGCCGAGTGTGTTGATCACCATCGATTAATAGTAAGTTTCGCAGATGACGTATCAGATTTCTACGGTCCCATGCTTGCTTTAAATTATATGTACCATTTAATTAGTTGCTGCCTTTTACTTCTAGAATGCTCGCAGAAGGAACCCGATGCCTTAGCACGTTACGGACCGCTTACAGTTATTATATTCGGTCAGTTGATTTCAGTGTCCGTTGTATTTGAAATCGTTGAAACGAAGGTAAAATCAACTAAATATGGGGCTAATTGCCTAACTTTAACTCTTTCCATGGTCCCATTCAGCAAGTGAGAATGAAACATAAGCTATTATTTAATAAGCTAATCTTTTTCCTAAATATGAAATTAATGAATGACCGCGCCCGAGTTACAGCTACACAATAAGCCCCCGAGTTCACTACTATACCTTGACTCCACTACAAAACTACCTTTAACCTAATAAGACAGACACATCGAAACCAATAAGTAATTATTTTATACCTTAAGTTCCTTCCCAACTAACATAGGTACTTGTGAAAAATGAAGTATTCCTTATAGCTCCAATGTCTATAATGGTGACATGATGGTGTCAAGATTAGGTACGAGTTCGAAAATAGCTCATGGTTTCATTTGCATGGAATAGCAGAATCCTAAGCAGGGCTTGATTTAATGCAGGGCAGACGGGGCTCTGGAAAAAATTGCTTTTGAGCTCGTAAAATTACATTTTTAAATAACTATTCTATAGTTCTCTATAGACTCCATAGTCTGCAGTGTATCATGATAGTATAGGTATTTGGTAAACCAATACAAAATAGAAATGAAATCAAAGAAAATTCTTCCTATGAGCGGTACTACAATATCAGAG
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xwchen24

金虫 (小有名气)

【答案】应助回帖

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感谢参与,应助指数 +1
常贺坦(myprayer代发): 金币+1, 赠人玫瑰,手有余香。分子生物期待你更多精彩 2013-08-25 12:13:12
1. 确定这个基因确实在你的模版中表达
2. 确定模版的质量,用别的引物P下看看
3. 改进PCR反应条件,你可以用下Touch-Down PCR,好使。
2楼2013-08-25 10:35:11
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