推荐回答(1个)
you should use some soft ware like clonemanager to design, to make your interested gene primers and the internal control gene primers have the similar annealing temperature can you give me the example that is similar to my requirements, please? i am looking forward your return.vs570588(站内联系TA)Originally posted by nininini at 2011-03-09 21:50:36:
做定量PCR的话目的序列大概在80~150bp就好了~~可以根据NCBI的序列查找一下保守序列,通过RNA序列分析挑选无二级结构的区段进行设计就好了~引物25个左右的bp吧,别太短,引物的特异性挺关键的 说真的降解现在我要处理的污染物微生物种类较多,我不是学生物出身的,搞水处理的,所以我也不知道怎样找保守序列,怎样算保守序列,我现在只能从NCBI中找到这种酶的序列,序列长度大约为1500个碱基左右。附图就是文献上设计引物时用到许多不同细菌表达这种酶的基因。你能给过我举个例子吗?
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