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8楼2011-06-08 15:57:23
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dream-fish

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silicare(金币+3): primer5 is ok 2011-06-07 18:30:50
693991425(金币+5): 2011-06-08 08:26:50
先克隆到基因之后,送测序公司测序。然后有软件把DNA序列直接转换成氨基酸序列就好了啊。我常用的就是primer 5.大部分生物序列分析软件都有这个功能。
2楼2011-06-07 16:47:49
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dream-fish

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693991425(金币+5): 2011-06-08 08:27:12
关于基因选取的问题就要看你做的这几个蛋白质是被人研究过吗?如果别人有研究过公布了序列就直接在NCBI上面去下载序列设计引物就好了。如果没有人研究过,那么就比较困难了,呵呵。
3楼2011-06-07 16:49:45
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693991425(金币+15): 2011-06-08 08:28:38
silicare(EPI+1): 2011-06-09 12:36:48
1. I assume proteins of interest are latest, namely other people have little research, because you are going to sequence them.
2. you should get coresponding mRNAs according to those proteins, not DNA.
3. How can you obtain the mRNAs of interest? you should acquire short amino acid sequence of those proteins through mass spectrometry. according to those short amino acid sequence, you can design degenerate oligonucleotide primers, then use those primers to clone genes (cDNA) of interest.
4. clone plasmid construction and sequencing them, or directly sequencing those DNA fragments.
5. you can acquire the amino acid sequence of those proteins according to those DNA sequences through triplet codon.
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4楼2011-06-07 22:32:33
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