I recently noticed and confirmed experimentally that the mouse 18S rRNA qPCR primers sold by a company that does not provide explicit primer sequence information actually specifically amplifies from the less abundant, immature 18S/5S/28S precursor RNA (primers span a maturation cleavage site). What effect might this have on use of these primers for qPCR normalization?
Hi David
This is really a matter of concern......and it may also happen in many other trasncripts where the primers designed in UTR regions may also amplify the precursor and not the mature transcript....I have also seen on such case in my lab...If the precursor is covnverted into the matur at the same efficeincy always...it should not be a problem...but if the conversion is affected by different conditions then it may affect the conclusions in qPCR assays.
Soone needs to be more carefull
byr
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