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Housekeeper Genes and Real-Time PCR

Housekeeper Genes L. Dev

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#1 Theo

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Posted 24 December 2016 - 02:34 PM

I am new to RT-PCR, Real-Time PCR,  as well as working with housekeeper genes. I analyze mice samples to determine if they have Lactate Dehydrogenase- elevating virus. I was given the task to find a house keeper gene that could help us ensure a negative result is truly a negative. I have searched literature and have found nothing published where this have been done. Can anyone led me in the direction of finding how to determine the best housekeeper genes and their primer sequences to accomplish the job I am trying to do? Thanks to anyone who can assist.



#2 bob1

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Posted 24 December 2016 - 07:37 PM

Basically all you need is some target that will work with all samples thereby ensuring that if it doesn't give signal for one sample you can assume that there was a problem with that sample, not that the test was returning a negative. Common genes to use for this include beta-actin and gapdh.
There are curated databases of primers for qPCR, but I can't remember the sites offhand. Try looking through the Nucleic Acids Research database issue.




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