Hi Varsha: It's not simply do one or two things to increase qPCR efficacy. We have learned from other investigators that kidney proximal tubule cells do not express gene coding alpha2-HS-glycoprotein (AHSG). We found however that it is not only detectable in qPCR but also in Western blotting (PMID: 27862254 ). We focused on three things: 1. obtain a high quality RNA (A260/280 ~2.0), 2. perform reverse transcription reaction using poly (dN)6 as primers, and 3. a good designed primer set for qPCR. Good luck.