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What housekeeper to use with HEK cells?


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#1 Stephp

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Posted 17 May 2013 - 03:39 AM

Hello everybody,

I am working with HEK293 cells and tried to check my RNA integration by using usual housekeepers (EF-1a, GAPDH, SDHD) but apparently they are not able to bind to this template, although in silico analyses revealed that they should. I tried them with quantitative PCR as well as with conventional RT-PCR with the same results.

I would be really happy for any suggestions.

Thanks a lot,

Best
Stephanie

#2 bob1

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Posted 17 May 2013 - 04:51 PM

Hmmm, those genes are all expressed by 293s, there is something wrong with either your PCR or your RNA extraction.

#3 Stephp

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Posted 18 May 2013 - 08:00 AM

Thanks, yes, that's what I thought. We checked the RNA (it was fine), transcription was done in parallel with other RNA samples that all worked well, and the house keepers gave good results in a qPCR using tissues that was run the same day...

Btw: are there possibly any differences between transfected vs. non-transfected HEK cells in expressing these genes?

Edited by Stephp, 18 May 2013 - 08:01 AM.


#4 bob1

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Posted 18 May 2013 - 05:44 PM

Potentially, you should test several housekeepers to find the ones (you should be using 2-3) that are most stable.




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