4 replies to this topic

[gyma]

Hi everyone, I tried to amplify a promoter region of my interested gene from genomic DNA of a cell line. Unfortunately, I didnot get a specific product. There might be problems elsewhere including the PCR conditions, primers and so on. But I want to make sure one thing first, is this promoter present in the genomic DNA of the cell line that I used? Although the gene is only expressed in specific tissues, I thought genomic DNA should be no big different among different tissues. That is why I didnt pay much attention to the template DNA and used the one that I have on hand. Am I wrong? Should I use the gene specific tissue for promoter amplification?

[bob1]

The promoter should be about the same in most cell lines from the same species, though there could be a number of mutations etc., that might have taken place, dependent on the cell line.

[gyma]

that means theoretically, I should be able to get the promoter sequence from the genomic DNA that I used, even if the gene is not expressed at all?

[bob1]

Yes.

[gyma]

bob1, on 14 July 2012 - 04:31 PM, said:

Yes.

thank you very much.