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Real time (CT IN NEGETIVE WELL AND DOBLE MELTING CURVE)


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#1 iman_r41

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Posted 02 September 2013 - 02:23 AM

hI

i have working on rbohD , after drought stress i cheked the exporession of this gene in my plant.

i have two problem

1. I got High CT in my negetive husekeeping gene (GAPDH) but after check in agarose i didnt see any thing. I am worry about my final result after normalosation with this house keeping gene.

2. sometimes double melting curve in my specific gene only in one accession. 

 

 



#2 iman_r41

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Posted 04 September 2013 - 05:44 PM


 



#3 Tabaluga

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Posted 09 September 2013 - 01:23 AM

To point 1. How high was your Ct ? If the Ct is more than 6 cycles higher than the sample with the lowest expression, it should be fine. You are aware that higher Ct stands for lower expression, I assume ?

To point 2 what do you mean by accession ? How does this look on the gel ? Primer dimers perhaps ?


Edited by Tabaluga, 09 September 2013 - 01:26 AM.

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.

 





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