I've been trying to amplify 3' splice variants of a hormone receptor gene, and I initially had great success. However, now I cannot repeat my results. The first 3-4 PCR's ran great, but now I only get a smear. I have replaced my primers because I was worried they were contaminated, but I still get the same smear. When I use different primers as a positive control, I get good clean amplification, so I think my cDNA is still OK. I need this specific reaction to work so I can amplify enough DNA to sequence the splice variants - any thoughts?

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