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  2. → Viewing Profile: Posts: AlexanderA

AlexanderA

Member Since 18 Nov 2010
Offline Last Active Jan 17 2013 02:30 AM
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Posts I've Made

In Topic: Multiple Bands in qPCR

10 January 2013 - 12:56 AM

Hi,

I just wanted to add a comment about my results. I did the qPCR again on another Cycler with rapid cycling (decreased annealing and elongation time), another mastermix, and annealing at 58°C.
The melting curve looks great now, everything is fine Posted Image

Best regards, and thanks again

In Topic: Multiple Bands in qPCR

21 December 2012 - 05:42 AM

Hi Trof,

thank you for your quick reply. I think I will try to decrease elongation time.
But just fort the understanding: What could the larger products be? The gDNA Amplicon should be approximately 16kb, which is quite unlikely to be amplified and obviously not on the gel.

Best regards Alex

In Topic: Reference RNA for qPCR

28 November 2012 - 10:51 AM

Hello Trof,

this sounds like a good Idea to me. Thank you very much.
I am wondering if there would be any benefit for the trial if I was performing a absolute quantitation with plasmids. I think to combine a relative quantitation with serial dilution for standard curve analysis from calibrator RNA and two endogenous reference genes should be fine to compare the gene expression levels between the two groups?

best regards,

Alex

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