发信人: glider (大苹果), 信区: Biology
标 题: Re: Sequence
发信站: Unknown Space - 未名空间 (Mon Oct 13 16:34:14 2003), 站内信件


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

【 在 windysea (雨瞳~像一个小孩只懂在你怀里坏) 的大作中提到: 】
: I recently got a big cDNA (coding sequence is 5.4 kb) from another lab they
: used in their publication,
: I sequenced 3 independent PCR products as well as the original one and in all
: cases, the sequence showed exactly the same 6 mismatches to the one in
: genebank. 4 cause single aa change, 2 are silent. What should I do now? It
: would be painful if I need to fix all of them in such a big cDNA.
: Is it possible that the sequence in genebank bears a couple of mistakes?


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