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Rotor-Gene 6000 data analysis


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#1 EmmaLin

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Posted 20 February 2014 - 02:41 AM

Maybe this is a silly question but here it goes.

 

I am using a Rotor-Gene 6000 with a commercial kit for Uraplasma qualitative detection.

 

this is actually a multiplex reaction: Green channel is Ureaplasma Parvum, Yellow channel is U.Uraliticum and Orange channel is internal control.

 

in my samples I observe the following. I get a signal (almost straight line) in the raw data Yellow channel, but when analysis is performed as specified by the manufacturer in the quantitation analysis channel no signal is seen or any line/curve. The software names these samples as NEG.

 

I am wondering if this is true negative or if this is a case of one probe in the multiplex failing. Is it normal to get a signal in the raw channel but no signal in the analysed data??

 

The IC works fine (nice curve on orange channel)

 

In the Green channel when a sample is negative I get a signal in the raw data BUT ALSO in the quantitation analysis data, below the threshhold of course.

 

Thank you



#2 phage434

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Posted 20 February 2014 - 04:51 PM

I think the only way to be certain is to find a positive sample, then do serial dilutions to establish sensitivity.






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