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[交流] 帮忙设计引物 产物100-500,超过九十分的60币

ATGTTCAAGGGGCTGAGCAAAGGCTCCCAGGGGAAGGGGTCCCCCAAGGGCTCCCCCGCCAAGGGGTCCCCCAAAGGCTCCCCCAGCAGGCACAGCCGGGCTGCCACCCAGGAGCTGGCCCTTCTCATCTCCCGCATGCAAGCCAACGCCGACCAGGTGGAGCGGGACATCCTGGAGACGCAGAAGAGGCTGCAGCAGGACCGGCTGAACAGTGAGCAGAGCCAGGCCCTGCAGCACCAGCAGGAGACGGGCCGCAGCCTGAAGGAGGCTGAGGTGCTGCTCAAGGACCTCTTCCTGGACGTGGACAAGGCCCGGCGGCTCAAGCACCCGCAGGCTGAGGAGATTGAGAAGGACATCAAGCAGCTGCACGAGCGGGTGACCCAGGAGTGTGCGGAGTACCGTGCCCTGTACGAGAAGATGGTGCTGCCCCCCGACGTGGGACCCAGGGTCGACTGGGCACGCGTGCTGGAGCAGAAACAGAAGCAGGTCTGCGCAGGCCAGTACGGGCCGGGCATGGCGGAGCTGGAGCAACAGATCGCCGAGCACAACATCCTGCAGAAGGAGATCGACGCCTATGGGCAGCAGCTGCGGAGCCTCGTGGGGCCGGATGCAGCCACCATCCGGAGCCAATACCGAGACCTACTGAAGGCGGCGTCGTGGCGCGGGCAGAGCCTGGGCAGCCTGTACACGCACCTCCAGGGCTGCACGCGGCAGCTGAGCGCCCTGGCTGAGCAGCAGCGCCGCATCCTGCAGCAGGACTGGAGCGACCTCATGGCCGACCCTGCGGGCGTGCGGCGGGAGTACGAGCACTTCAAGCAGCACGAGCTGCTGAGCCAGGAGCAGAGCGTGAACCAGCTGGAGGACGACGGCGAGCGCATGGTGGAGCTGCGGCACCCCGCGGTGGGGCCCATCCAGGCCCACCAGGAGGCCCTGAAGATGGAGTGGCAGAACTTCCTGAACCTGTGTATCTGCCAGGAGACCCAGCTGCAGCACGTGGAGGACTACCGCCGGTTCCAGGAAGAGGCCGACTCAGTCAGCCAGACCCTGGCGAAGCTCAACTCCAACTTGGATGCCAAGTACAGCCCTGCACCTGGGGGCCCCCCTGGCGCCCCCACAGAGCTGCTGCAACAGCTGGAGGCAGAGGAAAAACGGCTGGCCGTCACCGAGAGGGCCACTGGGGACCTGCAGCGGCGAAGCCGGGATGTGGCCCCTCTGCCACAGCGAAGAAACCCCCCTCAGCAGCCCCTGCACGTGGACAGCATCTGCGACTGGGACTCAGGAGAAGTGCAGCTGCTGCAGGGTGAGCGGTATAAGCTGGTAGATAACACTGACCCGCACGCCTGGGTCGTGCAGGGCCCTGGCGGGGAGACCAAGCGTGCTCCCGCCGCCTGCTTCTGCATCCCAGCACCAGACCCTGATGCTGTGGCCAGGGCCTCCCGGCTGGCCTCAGAGCTGCAGGCCCTGAAGCAGAAATTGGCCACAGTCCAGAGCCGCCTGAAGGCCAGTGCTGTGGAGTCTCTTCGGCCCAGCCAGCAGGCTCCATCTGGCTCAGACCTGGCCAACCCACAGGCCCAGAAGCTCCTGACACAGATGACCCGGCTGGATGGAGACCTGGGACAGATAGAGAGGCAGGTGCTGGCCTGGGCGCGGGCCCCGCTGAGCCGCCCCACACCCTTGGAGGACTTGGAGGGCCGCATCCACAGCCATGAGGGCACAGCCCAGCGCCTGCAGAGCCTGGGAACGGAGAAGGAGACAGCCCAGAAGGAGTGCGAGGCGTTTCTGTCCACGCGGCCCGTGGGCCCCGCTGCCCTGCAGCTGCCCGTAGCCCTCAACAGCGTGAAGAACAAGTTCAGTGACGTGCAGGTTCTGTGCAGCCTCTACGGGGAGAAAGCCAAGGCTGCCCTGGATCTGGAGCGGCAGATCCAGGATGCGGACAGGGTCATCCGAGGCTTCGAGGCCACCCTGGTGCAGGAGGCCCCCATCCCTGCTGAACCGGGGGCTCTGCAGGAGAGGGTCAGCGAGCTGCAGCGCCAGCGGAGGGAGCTGCTGGAACAGCAGACCTGCGTGCTGCGGCTACACCGCGCGCTGAAGGCCTCGGAGCACGCATGCGCTGCCCTGCAGAACAACTTCCAGGAGTTCTGCCAAGACCTGCCTCGCCAGCAGCGCCAGGTGCGAGCCCTCACCGACCGCTACCACGCCGTAGGGGACCAGCTGGACCTGCGGGAGAAGGTGGTGCAGGATGCCGCCCTCACCTACCAGCAGTTCAAGAACTGCAAGGATAACCTGAGCTCCTGGCTGGAGCACCTGCCCCGCAGCCAGGTGCGGCCCAGCGACGGCCCCAGCCAGATCGCCTACAAGCTGCAGGCGCAGAAGAGGCTGACGCAGGAGATCCAGAGCCGAGAGCGGGACAGGGCCACAGCATCCCACCTCTCCCAGGCCCTGCAGGCAGCGCTCCAGGACTATGAGCTCCAGGCAGACACCTACCGCTGCTCTTTGGAGCCCACCCTGGCAGTGTCAGCCCCCAAGAGACCCCGAGTGGCTCCCCTGCAAGAGAGCATCCAAGCCCAGGAGAAGAACCTTGCAAAGGCCTATACTGAGGTTGCAGCAGCACAGCAGCAGCTGCTCCAGCAGCTGGAGTTTGCTAGAAAAATGCTGGAGAAGAAGGAGCTCAGTGAGGACATCCGAAGGACCCATGATGCAAAGCAGGGCTCCGAGAGCCCTGCCCAAGCAGGGAGAGAGTCAGAGGCCCTGAAGGCCCAGCTGGAAGAGGAGAGGAAGCGGGTGGCCCGGGTGCAGCATGAGCTGGAGGCGCAGAGGAGCCAACTGCTGCAGCTGAGGACCCAGCGGCCCTTGGAGAGGCTGGAGGAGAAGGAAGTGGTAGAGTTCTACCGGGACCCCCAGCTGGAGGGCAGCCTGTCCAGGGTGAAGGCCCAGGTGGAGGAGGAGGGCAAGCGGCGGGCTGGCCTGCAGGCAGACCTGGAAGTGGCAGCCCAGAAGGTCGTGCAGCTGGAAAGCAAGAGGAAGACCATGCAGCCTCATCTGCTGACCAAGGAGGTCACCCAGGTGGAGAGGGACCCCGGCCTGGACAGCCAGGCGGCCCAGCTCAGGATCCAGATCCAGCAGCTCCGCGGGGAGGATGCCGTCATCTCGGCCCGGCTGGAAGGGCTGAAGAAGGAGCTACTGGCCCTTGAGAAGAGGGAGGTGGACGTGAAGGAGAAGGTCGTGGTGAAAGAGGTAGTCAAGGTGGAGAAGAATCTGGAAATGGTCAAGGCAGCCCAGGCTCTGAGGCTGCAGATGGAGGAGGATGCTGCGCGGAGGAAGCAGGCGGAGGAGGCTGTGGCCAAGCTACAGGCTCGCATCGAAGACCTGGAGCGGGCTATCAGCTCGGTGGAGCCCAAGGTCATCGTGAAGGAGGTGAAGAAGGTGGAGCAGGACCCAGGGCTCCTCCAGGAGTCCTCCAGGCTGAGGAGCCTCCTCGAGGAGGAGAGGACCAAGAACGCGACGCTGGCCAGGGAGCTGAGCGACCTGCACAGCAAGTACAGCGTGGTGGAGAAGCAGAGGCCCAAAGTGCAGCTCCAGGAGCGCGTCCACGAGATCTTCCAGGTGGATCCGGAGACAGAGCAGGAGATCACTCGGCTCAAGGCCAAGCTGCAGGAGATGGCGGGCAAGAGGAGCGGTGTGGAGAAGGAGGTGGAGAAGCTGCTGCCCGACCTGGAGGTCCTGCGGGCCCAGAAGCCCACGGTGGAGTACAAGGAGGTGACCCAGGAGGTGGTGAGGCATGAGAGGAGCCCCGAGGTGCTGCGTGAGATCGACCGCCTGAAGGCTCAGCTCAACGAGCTCGTCAACAGCCACGGGCGCTCCCAGGAGCAGCTCATCCGCCTGCAGGGTGAGCGCGACGAGTGGAGGCGCGAGCGGGCCAAGGTGGAGACCAAGACGGTGAGCAAGGAGGTGGTGCGCCACGAGAAGGACCCGGTGCTGGAGAAAGAAGCAGAGCGGCTCCGCCAGGAGGTGCGGGAGGCGGCCCAGAAGAGGCGGGCCGCGGAGGACGCGGTGTACGAGCTGCAGAGCAAGCGCCTGCTGCTGGAGAGGAGGAAGCCCGAGGAGAAGGTGGTGGTGCAGGAGGTGGTGGTCACCCAGAAGGACCCGAAGCTGCGCGAGGAGCACAGCCGGCTGAGCGGGAGCCTGGATGAGGAGGTGGGCCGGCGGCGCCAGCTAGAGCTTGAGGTGCAGCAGCTGCGGGCCGGCGTGGAGGAGCAGGAGGGCCTGCTCAGCTTCCAGGAGGACCGCAGCAAGAAGCTGGCCGTGGAGAGGGAGCTGCGGCAGCTGACCTTGAGGATCCAGGAGCTCGAGAAGCGGCCTCCCACGGTGCAGGAGAAGATCATCATGGAGGAAGTGGTCAAGCTGGAGAAGGACCCGGACCTGGAGAAGTCCACGGAAGCCCTGCGGTGGGACCTGGACCAGGAGAAGACCCAGGTAACCGAGCTGAATCGGGAGTGCAAGAACCTGCAGGTCCAGATTGACGTCCTCCAGAAAGCCAAATCGCAGGAGAAGACCATCTACAAGGAAGTGATCCGGGTGCAGAAGGACCGCGTCCTGGAAGATGAGCGGGCCCGCGTGTGGGAGATGCTCAACAGGGAGCGCACGGCCCGGCAGGCCCGGGAGGAGGAGGCACGGCGCCTGCGGGAGCGCATTGACCGGGCCGAGACGCTGGGGAGAACCTGGTCCCGGGAGGAGTCCGAGCTGCAGAGGGCCCGGGACCAGGCCGACCAGGAGTGTGGGCGGCTGCAGCAGGAGCTGCGGGCTCTGGAGAGGCAGAAGCAGCAGCAGACACTGCAGCTGCAGGAGGAGTCGAAGCTGCTCAGCCAGAAGACGGAGAGCGAGCGACAGAAGGCGGCCCAGCGGGGCCAGGAGCTCTCGCGGCTGGAGGCGGCCATCCTCCGCGAGAAGGACCAGATCTACGAGAAGGAGCGGACGCTCCGGGACCTCCACGCCAAGGTGAGCCGGGAGGAGCTCAGCCAGGAGACCCAGACGCGAGAGACCAACCTTTCCACCAAGATCTCCATCCTGGAACCCGAGACGGGGAAGGACATGTCCCCATACGAGGCCTACAAGAGGGGCATCATCGACAGGGGCCAGTACTTGCAGCTGCAGGAGCTCGAGTGTGACTGGGAGGAGGTCACCACCTCGGGGCCCTGTGGGGAGGAGTCTGTGCTCCTGGACCGCAAGAGCGGGAAGCAGTACTCCATCGAGGCCGCCCTCCGCTGCCGGCGCATCTCTAAGGAGGAGTACCATCTGTACAAGGACGGCCACCTGCCCATCTCCGAGTTTGCGCTGCTTGTAGCTGGGGAGACCAAGCCAAGCTCCTCACTCTCCATCGGCTCTATCATCTCCAAGTCCCCGCTCGCCTCCCCGGCCCCCCAGAGCACCAGTTTCTTCTCTCCCAGCTTCTCTCTCGGGCTCGGTGATGACAGCTTCCCTATCGCCGGGATCTATGACACAACCACAGACAACAAGTGCAGCATCAAGACGGCCGTGGCCAAGAACATGCTGGACCCCATCACTGGGCAGAAGCTACTGGAGGCCCAGGCGGCCACAGGGGGCATCGTGGACCTGCTCAGCCGTGAGCGCTACTCTGTGCACAAGGCGATGGAGAGGGGCCTGATCGAGAACACCTCCACACAGAGGCTGCTTAACGCCCAGAAGGCCTTCACCGGCATCGAGGACCCCGTCACCAAGAAGAGGCTCTCGGTGGGCGAGGCCGTCCAGAAGGGCTGGATGCCCCGGGAGAGCGTGCTCCCACACCTGCAGGTGCAGCACCTGACCGGGGGGCTCATCGACCCCAAGAGGACAGGCCGCATCCCCATCCAGCAGGCCCTCCTCTCCGGGATGATCAGTGAAGAGCTGGCCCAGCTCCTGCAGGACGAGTCCAGCTACGAGAAGGATTTGACAGACCCCATCTCCAAGGAACGGCTGAGCTACAAGGAGGCCATGGGCCGCTGCCGCAAAGACCCCCTGAGCGGCCTGCTGCTCCTGCCAGCGGCACTGGAGGGGTACCGCTGCTACCGCTCCGCCTCCCCCACCGTCCCGCGCTCCCTTCGCTGA
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已阅   回复此楼   关注TA 给TA发消息 送TA红花 TA的回帖

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1949stone(金币+5,VIP+0):够热情 11-25 10:01
fengwenbin(金币+5,VIP+0):谢谢,我是做rt-p,struggle很久了,循环中30度要6‘30?望详细指导谢谢 11-25 22:52
你的这个给基因好大,引物最好长一些,以我4年经验,引物为下:

正向:5' atgttcaaggggctgagcaaaggctcccagg 3'

反向:5' tcagcgaagggagcgcgggacggtggg 3'

建议用LA TAQ 酶做PCR,条件:
94度 5分钟

94度 30秒
72度 6分钟30秒
25个循环

72度 10分钟

如果你是做TR-PCR,做最分做两步
如果以上条件不行,也设计4条引物,先PCR得到两条片段,在OVER LAPPING PCR
6楼2009-11-25 09:50:19
已阅   回复此楼   关注TA 给TA发消息 送TA红花 TA的回帖
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