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NWAMN

新虫 (初入文坛)

[求助] 如何拼接两段序列,急! 已有2人参与

毕业实验我想要扩一个基因的全长,但是这个基因的3端的序列未知,所以我的得先测出未知序列。下面是我做实验测序的3端的一段未知序列:
GATTATCCTGGTGAAAATGCAAGTCTCAAAAAGAAAAGAAAAAGAAGATGATAAAACTTCTAAAAATAAAAAAGACGTGAGTGTTAGAATGAGCAC
ACTTTCATATGATGACAATTCTTCTCTCTCGACGACCACGCGTGCCCTATAGT
后面想将它和前面已知序列拼接,已知5端序列为:
ATGCCTCCATCACAATTATTCAAAGTTCATGGAGAATTCTGTGCAGCATATCCATGGGAAGTTATTGTGGCAACATTAACATTAGCTGTATGTATGTTAACAGTTGACCAACAACAACATTCAACGATACCACCTAAACCTATAGTACGAAACTGTGCTGGTTGTCTACACGAGGCAGAATATAATGCTGCTGACGTGATTGTTATGACTTTAATAAGATGCTTAGCTGTACTTTACAGTTATTATCAATTCTGCAATCTACAAAGATTAGGATCAAAATATATTTTGGGTATTGCCGGTTTATTCACAGTTTTCTCCAGTTTCATATTCACAACAACTGTTTTAAATCTGTTACGTATCGATATGTCCGATTTAAAGGATGCATTATTTTTCTTTCTACTATTAATAGATTTATCAAAAGCGGCAATGTTAGCACAATCGGCATTAAGCGCATCAAACCAAGATGAAGTTAAAAGTAATATTGCGAGAGGTATGGCAATTTTAGGTCCCACAATAACATTGGATACAATTGTTGAAACTTTAGTTATTGGTGTCGGTACGCTTTCCGGTGTCCAACGTCTAGAGATGCTCTCATATTTTGCCTGTCTTTCAGTTTTAGTTAATTACATTGTCTTCATGACATTCTATCCGGCTTGTTTATCATTAATATTAGAGTTATCTCGCACTACAAATATTTACGGTAAGAAGCAATCGTTAATATCACGAGCATGGAAAGAAGAGGACCACAAATCAAATCCTGTTGTACAAAGAGTTAAATTGATTATGTCAGCTGGTTTGATGTTAGTACATGCTCGTAGTCGTTGGCCTTTCAAAGCTGATGAAGTTGATAGTATTGGACCTTTAGTTGTTGAACAGCATATGGGACAAAATCGTACAGAAGATACAGCTTTACACGAATATATGATGAAATGGGTTACTGTAAGTGCTGATCACATCGTGATTTTAATTCTACTATTAGCATTAGTGGTGAAGTTTATATTTTTCGAAAATAAAGACGATTTAGCACTACAATTGCGCGTTCATATTAGTGAAAGTAACGAAAATAAAAAGATCGAACCAACGAAAAAAATGCCATTTTTGAAACCGCAAACATTCTTTCTAGAAGACAAAGATATAGCGGAAAATTCCAAAGCAACTGAATTCGAAGAAAAAGGAGTACAAACGGAAAAATTAAAACACGACTCAAATACAGAAATAACTCAAATTGAAGATGATACAAATAAACAAGAGATTTGTAGAAGTTTAGATGAATGCCGACGTATTTACAATGATACAGATTTAGGTGGTACATATTTAACTGATGATGAAGTAATATCACTTGTGCATAGTAAAGATATAGCAGCATATCAAATAGAGAAAGCTGTTGAAGATCCAGTACGTGGTGTTGGGATCCGGCGTAAGATTGTTGGCCATGAAGGCAATTTACTAAAAGCACTTACCGATTTACCATATAAACAATATGATTATAGTAAAGTTATGGGGGCTTGTTGTGAAAATGTTATCGGTTATATGCCAGTACCAGTTGGTATAGCTGGACCATTAAATTTGGACGGTCGTCCTGTTTATATACCAATGGCAACAACAGAAGGATGTCTTGTAGCCAGTACAAATCGTGGTTGTCGGGCTTTGAATGAATGTGGTATTATTAGTTGTATTGTTGCTGATGGTATGACTAGAGGACCAGTTGTTCGATTTCCATCAATAAAAAAAGCTGGTGAAGCTATGGCTTGGATGAAATCAAACGATAATTTCGCTTTAATTAAACAACATTTCGATGCTAGTAGTCGATTTGCACGTTTATCAAAATTATTAATTAAAATAGCCGGACGTCATCTGTTTATTCGTTTCGAAGCGAAAACTGGAGATGCTATGGGTATGAATATGTTATCAAAAGGGACTGAATTATCATTAAAATACATACAAAAAACACATTTTCCTGAAATGGAGATACTCAGTTTAAGTGGTAATTTCTGTACTGACAAGAAACCGGCTGCTGTTAACTGGATCGAAGGTCGTGGTAAATCGGTAGTTTGTGAGGCTATAGTTCCAGCAAAGATAGTAACAAAAATATTAAAAACGAGCGCATCTGCTATTGTTGATGTTAATAATTCAAAGAATATGATTGGTTCAGCTGTTGCCGGCAGTATAGGTGGTTTTAATGCACATGCGGCTAATATAGTTACAGCAATTTATATAGCAACAGGCCAAGATCCAGCACAAAATATTGGTAGTAGTAATTGTATGACATTAATGGAAACATGGGGTGAAACTGGTGAAGATTTATACATATCATGTACAATGCCATCTATTGAAATTGGTACAATTGGTGGTGGTACAATATTGCCAGCACAGGCTTCTTGTTTAGAAATTTTGGGTGTTAAAGGGCCACATCAAGATTATCCTGGTGAAAATGCAAGTCAATTAGCACGTATA GTTTGTGCAACTGTATTGGCTGGTGAATTATCATTGATGGCCGCTTTAACAGCTGGACATTTG

但是我一直拼接不上,用DNAMAN拼接的,不知手工如何拼接呢?或者用DNAMAN不知哪个地方有问题?
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NWAMN

新虫 (初入文坛)

引用回帖:
2楼: Originally posted by stone2239 at 2015-06-01 23:00:45
不是拼接的问题,是你扩出的序列不对,

怎么判断啊,但是能找到引物啊
4楼2015-06-02 18:23:08
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stone2239

铁杆木虫 (著名写手)

【答案】应助回帖

感谢参与,应助指数 +1
不是拼接的问题,是你扩出的序列不对,
2楼2015-06-01 23:00:45
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xmcliulei

银虫 (小有名气)

【答案】应助回帖

感谢参与,应助指数 +1
同学你基因全长多长啊?这两段你确定有重叠区么?
3楼2015-06-01 23:25:40
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NWAMN

新虫 (初入文坛)

引用回帖:
3楼: Originally posted by xmcliulei at 2015-06-01 23:25:40
同学你基因全长多长啊?这两段你确定有重叠区么?

由于3端是未知序列,所以不知道有多长,有重叠区,但不是太长
5楼2015-06-02 18:24:17
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