I exposed fish as embryos to 10, 50 and 200ppb arsenic and now at 28 weeks I am utilizing qpcr to look at a few genes. I have never had this problem on all other plates but when i run 18s for my female liver samples I see a drastic increase in Ct values for the 50 and 200 ppb liver cDNA. Does anyone know why? I have run several plates and even ran plates with newly made cDNA but I am still seeing the same issue.
Hi
surprising, did you test QC of your RNA just to ensure it's still there in enough quality?
fred
Yes we checked this 3 times and every time the ABS 260/280 was greater than 1.7 with an average of 1.9 for all samples and the concentrations were all high enough to proceed. It is so strange and we have not been able to figure it out as to why we only see this at 28 week old female killifish liver.
- Badges
- Science method