zoyd81

Hi

We are working on gene expression of two opioid receptors in human skin: OPRM1 and OPRK1 using universal probe library, UPL probes from Roche company. We do not have any problems with OPRK1 becouse it.s gowing fine on all probes, but problems starts with OPRM1. We detect by Probe Finder three different primer pairs, which react with constitutive regions for OPRM1. In the begining we make real-time but using Sybr Green. For positive control we use cDNA from human brain. All primers react fine, in the brain and in the skin. Melting curve also look good.Sequencioning results give us the same sequences in the brain, skin as reference sequence in NCBI. The same reaction we do on the same material: brain and skin cDNA but using UPL probes. All primers and UPL probes detect expression of OPRM1 in the brain, but in the skin not !!!
We have the same problem with OPRM1 when we use ready to use gene expression assays from Applied Biosystem. In brain everything was ok, but in the skin we do not have any expression.
We also make this experiments using very good cDNA material from the keratinocyte cultures. OPRM1 work with sybr but not with UPL probes.

Please HELP