Hi,
Apologize in advance for the incompetence - I am new to that system. I am also new to the field and to the cell work - that project is different from what I have been doing before.
I am planning to perform some experiments to figure out if my piwil protein(s) is(are) expressed in some human cell lines. However I definitely need a positive control for that (where these protein(s) will constantly be expressed). I am thinking about some testicular cell line (they say that in mammals piwi genes are expressed specifically in testis) however I don't know what could be the best solution for that? (like if there any widely used cells for such purposes?)
Hope I made it clear more or less, I am thanking you in advance.
Thank you, Bruno.
Well, there's an article (for ex. this one http://www.ncbi.nlm.nih.gov/pubmed/12906857) where people are using the multitissue cDNA panels, showing that "all 4 members of the PIWI subfamily are expressed mainly in testis" and there's a figure (Fig 4) showing the signal on "testis". So I was wondering what kind of cells I may use as a positive control for the RT-PCR I would like to do...
After all, any useful information would be useful! So, please let me know if you find something..
Ok, thank you, Bruno.
So what in your opinion could be the "versatile" human testis cells that I might like to find? (Perhaps ordering of the cDNA would be the easiest soultion however since I am completely new to the rt-pcr technique I'd like to prepare the cDNA by myself).
Does, for example, that cell line from ATCC http://www.atcc.org/Products/All/CRL-2073.aspx looks fine for you for these piwil positive control purposes that I require?
Thank you!
For positive controls (in vivo) we use testis tissue samples from mouse. But if you are specifically looking for human PIWI, ordering RNA made from human testis will be better option.
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