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low raw fluorescence


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#1 sponge_nuts

sponge_nuts

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Posted 09 September 2009 - 05:31 PM

Hi guys,
I'd like to know your opinion about the two amplification plot I've attached.
It's the same gene, same template, same primers, same amplification conditions and machine settings (Corbett 3000). I'm using Sybr GreenER (Invitrogen) but the raw fluorescence is very low in 2009 run vs. the 2008.
the technical support told me that the 2009 curve is good, but it doesn't look like an exponential amplification and it has nothing to do with the 2008. I'm having the same problem with other genes that are being amplified with a rising straight line and a very low fluorescence. I tried to use freshly synthesized cDNA and fresh reagent but it is even worse..
Any suggestion?

Cheers

Attached Thumbnails

  • 2008.jpg
  • 2009.jpg





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