We plan to monitor the expression of certain virulence genes during acid shock treatment, We would like to know if the housekeeping genes used will also be affetcted by this exposure?
This would be an empirical evaluation (in agreement with Sudip Das' comment above). Be sure to test and find at least 3 reference genes (HKGs) unaffected by your acid shock treatment. You will not know what genes can serve as stable reference genes for your particular system until you test them. It is not uncommon to run more tests to find good ref. genes than you do for your genes of interest.
Also, perhaps Google acid shock stable ref. genes (or house-keeping genes) for RT-PCR/qPCR/RT-qPCR in bacteria.
This would be an empirical evaluation (in agreement with Sudip Das' comment above). Be sure to test and find at least 3 reference genes (HKGs) unaffected by your acid shock treatment. You will not know what genes can serve as stable reference genes for your particular system until you test them. It is not uncommon to run more tests to find good ref. genes than you do for your genes of interest.
Also, perhaps Google acid shock stable ref. genes (or house-keeping genes) for RT-PCR/qPCR/RT-qPCR in bacteria.
Hello,
Agree with both. Before commencing any RT-PCR experiments, you should check the basal expressions of minimum of 3 HKGs by usual qPCR or semi-qPCR.
Regards,
Sanjay
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