g*r
2 楼
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
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
J*S
3 楼
惭愧，下线，不灌水了
g*r
4 楼
Oh, I omit an E
it is I.M.A.G.E. -you can check the clone out in ncbi, to see if there is information
on this cDNA/entry, if there is something like an image number, or atcc number
then you can find the clone from atcc website, order it, about $50~ bacteria ...
it is I.M.A.G.E. -you can check the clone out in ncbi, to see if there is information
on this cDNA/entry, if there is something like an image number, or atcc number
then you can find the clone from atcc website, order it, about $50~ bacteria ...
r*o
5 楼
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
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
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