internal standard for semi-quantitative pcr

a.boasso a.boasso at tiscalinet.it
Mon Dec 11 14:58:43 EST 2000


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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