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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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Sybr green
Posted by: xuan (IP Hidden, New member, 1)
Date: November 9, 2005 08:11PM
Hi-I was wondering if anyone can help me with this problem. Sometimes when I did RT-PCR using SYBR green, I got this wavy curve that was not as smooth as it should be but otherwise normal. I'm not sure if I should be concerned about this wavy thing. Does anyone also get wavy curves and does anyone know what's the cause? Thanks!
Re: Sybr green
Posted by: Fabian (IP Hidden, New member, 1)
Date: December 2, 2005 09:19AM
Some questions first to get an idea whether maybe I have some idea or not ;)
1. How much SybrGreen are you using? 2. It is a onestep or twostep RT-PCR? 3. If it is a twostep RT-PCR how much cDNA you are using and do you use hexamer/specific/Oligo(dT) Primer? 4. Where is the RNA from? Which kind of isolation you are using? 5. Ocurs this problems with the same Primers/Templates/PCR.conditions?
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