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how to hasten real-time PCR amplifications


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#1 yobou

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Posted 04 March 2014 - 11:23 PM

hello there

when I perform real-time PCR for different genes in human cancer cells, I usually got a base-line until about 25 cycles then linear amplifications starts. I use the following conditions:

1) cDNA concentration/well ~29 nanograms

2) forward or reverse primers concentration : 350 nM (final)

 

How can I shorten the length of the base-line and get amplifications earlier after about 15 cycles?

thanks in advance



#2 Tabaluga

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Posted 04 March 2014 - 11:40 PM

In principle with a higher cDNA amount

Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 


#3 vetticus3

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Posted 05 March 2014 - 07:31 AM

seconding the more cDNA.






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