发信人: royluo ( 罗马情结), 信区: Biology
标 题: Re: Sequence
发信站: Unknown Space - 未名空间 (Mon Oct 13 18:22:53 2003), 站内信件


IMAGE clones估计是没戏的。IMAGE 是Integrated Molecular
Analysis of Genomes and their Expression 的缩写。网址http://
image.llnl.gov. 实际上是一个EST cDNA clones library. 你可以
在invitrogen订购，http://clones.invitrogen.com/
收得倒挺全，但是序列可能会有问题，在3'端常常有错误，出现stop
codon什么的。你这个基因这么大，很难保证有完全正确的cDNA.
当然你也可以选择一些测过全长，甚至可以直接表达的clone库，
同样是http://clones.invitrogen.com. 但是这种库clone十分有限，
你要的基因不一定在里面。另外日本有个免费的克隆库，
http://www.kazusa.or.jp/huge/ , 你可以去找找。
另外，实际序列与网上序列库序列有微小的不一致，其实一点也不
奇怪。我就遇到过这种情况。建议你先问问给你clone的实验室，看
他们的意见如何�



【 在 windysea (雨瞳~像一个小孩只懂在你怀里坏) 的大作中提到: 】
: as I said, they are same mismatches on both pcr products and the original one.
: I was using pfuUltra HF in the kit since I was doing site directed mutagenesis
: instead of cloning. pcr should not be the problem, since it only introduced
: the
: designed mutation for mutagenesis. Other mismatches are also present in the
: original one.
: Can you give me the information for IMAG? anyway, I doubt if it has it since
: this one is big and relatively novel.
: 【 在 glider (大苹果) 的大作中提到: 】
: : it is possible the gene in the bank has mistakes
: : but what pcr enzyme did you use, since taq tends to give errors
: : besides, you may order the clone from IMAG directly, instead of cloning
: : by yourself, if you are sure the clone is the one that you are going to use
: : I guess it is cheaper to buy the clone, than do the whole cloning/sequencing
: by your own
: they
: all


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