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源子不二

新虫 (初入文坛)

[求助] 恳请大神帮助设计race引物,5个金币太少,但真是全部家当了 已有1人参与

发帖求助时,才发现全部家当只有5个金币,我知道太少了。。。请大神别嫌弃,先谢谢了(万一大神在西安,我请你吃海底捞抵金币可以吗)。我用自己设计的两对引物做了好久都做不出来,用的是smart试剂盒。闷头研究了几天想再设计引物可是一点信心都没有,打电话给生工,生工不给设计race引物。希望小木虫上热心大神帮助。非常不好意思我研究了下不会上传word。。。只能麻烦大神亲自ncbi搜一下。或者请留下邮箱,我给发过去。
基因名称:Sitophilus zeamais cytochrome oxidase subunit I (COI) gene, partial cds; mitochondrial gene for mitochondrial product   
GenBank: AY131100.1   
ACCESSION   AY131100
用的试剂盒里的引物有:
10X Universal Primer A Mix (UPM)
Long (0.4 μM): 5'–CTAATACGACTCACTATAGGGCAAGCAGTGGTATCAACGCAGAGT–3'
Short (2 μM): 5'–CTAATACGACTCACTATAGGGC–3'
Nested Universal Primer A (NUP; 10 μM) 5'–AAGCAGTGGTATCAACGCAGAGT–3'Control Reagents

再次感谢,希望虫友大神能给我一道光。

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源子不二

新虫 (初入文坛)

引用回帖:
3楼: Originally posted by mlbiosci at 2015-04-20 18:12:31
先比对序列选取同源区,设计引物,分别做3“  5"race

谢谢~!

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4楼2015-04-21 09:08:32
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源子不二

新虫 (初入文坛)

各位大神,序列在这里。
>gi|33383489|gb|AY131100.1| Sitophilus zeamais cytochrome oxidase subunit I (COI) gene, partial cds; mitochondrial gene for mitochondrial product
TCTCTACTAACCATAAAGATATTGGAACATTATACTTTATTTTTGGAGCATGATCAGGAATAGTGGGAAC
CTCATTGAGTTTACTAATTCGGGCTGAATTAGGGAATCCTGGATCATTAATTGGAAATGATCAAATTTAC
AATACTATTGTTACAGCTCACGCATTCATTATAATTTTCTTTATAGTGATACCTATTATAATTGGAGGTT
TCGGAAACTGATTAGTCCCACTAATACTAGGAGCCCCAGATATAGCATTCCCACGATTAAACAATATAAG
GTTCTGATTACTCCCTCCATCATTAATTCTTTTATTAATAAGAAGATTCATTGAAAAAGGTGCCGGAACA
GGATGAACTGTTTATCCCCCATTATCCTCAAATATTGCACACGAAGGAGCCTCTGTTGATTTAGCAATTT
TTAGTCTTCATATAGCAGGTATTTCATCTATTCTTGGAGCTATTAATTTTATTTCTACTACTTATAATAT
ACGCCCCTCCGGTATATTATCAGATCGAATAACTTTATTTATTTGAGCAGTTAGAATTACAGCCATTCTT
CTTTTACTTAGATTACCTGTTTTAGCTGGAGCAATCACTATGCTTCTTACTGATCGTAATATCAATACCT
CTTTTTTTGATCCTGCTGGAGGCGGAGACCCTATTCTCTATCAACACTTATTTTGATTCTTCGGACACCC
AGAAGTTTACATTTTAATTCTCCCAGGATTTGGNATAATCTCTCATATTATTAGTTACGAAAGAGGAAAA
AAAGAAACTTTCGGAGTATTAGGAATAATCTATGCAATAATCGCAATTGGTTTATTAGGATTCGTAGTTT
GAGCTCATCATATATTTACAGTAGGAATAGACGTTGATACACGAGCATACTTTACATCAGCTACTATAAT
TATTGCAGTCCCTACAGGAATTAAAATTTTTAGATGATTAGCAACTTTTCATGGTGCACGAATTACCTAT
TCTCCTACATCCCTTTGATCTTTAGGATTTATTTTCCTTTTTACTATCGGGGGATTAACAGGAGTAGTTT
TGGCCAATTCATCTATTGACATTATTCTTCATGATACGTACTATGTTGTA

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2楼2015-04-20 15:21:59
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mlbiosci

新虫 (正式写手)

【答案】应助回帖

感谢参与,应助指数 +1
先比对序列选取同源区,设计引物,分别做3“  5"race

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3楼2015-04-20 18:12:31
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