internal standard for semi-quantitative pcr

Mark Haynes Mark.Haynes at Mail.TJU.Edu
Tue Dec 12 18:22:07 EST 2000


gapdh is another common sequence that is used as a ubiquitous signal
but I don't know if its applicable for you

"a.boasso" wrote:

> We are using a semi-quantitative pcr assay to evaluate the expression level
> of genes involved in the immune system functions (like citokynes, receptors,
> co-stimulation molecules), in hiv+ individuals or aids patients.
> In this assay we perform a pcr reaction with two sets of primers:
> 1. a pair of primers for the target mRNA amplification (citokynes, receptors
> or co-stimulation molecules)
> 2. a pair of prime for the beta-actin mRNA, which will be the internal
> standard
>
> The beta-actin mRNA concentration is determined in a previous step, with a
> specific competitive pcr assay, with a synthetic competitor, used in
> different reactions at different concentrations.
>
> Some critics have been moved against the use of beta-actin as an internal
> standard, as its expression seems to change widely in pathologic cells.
>
> Is anybody able to give me some hints or information about the use of some
> other internal standard genes in cells from hiv infected patients?
>
> Thank's






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