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Limit of detection

Limit of detection - Real-Time PCR and Quantitative PCR Forum

Limit of detection - Real-Time PCR and Quantitative PCR Forum


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  #1  
Old 08-01-2011, 05:26 PM
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Default Limit of detection



Hello,

I need to decrease the limit of detection in my real time PCR. In the process of standarization i have to amplify 19/20 samples in the SmartCycler. However, I always have more than 3 negatives!

I would like someone to tell me some advices please!

Thannnks
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Old 08-01-2011, 05:26 PM
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Default Limit of detection

Hello,

I need to decrease the limit of detection in my real time PCR. In the process of standarization i have to amplify 19/20 samples in the SmartCycler. However, I always have more than 3 negatives!

I would like someone to tell me some advices please!

Thannnks
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  #3  
Old 08-01-2011, 05:27 PM
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Default Limit of detection

Hello,

I need to decrease the limit of detection in my real time PCR. In the process of standarization i have to amplify 19/20 samples in the SmartCycler. However, I always have more than 3 negatives!

I would like someone to tell me some advices please!

Thannnks
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  #4  
Old 09-28-2011, 10:44 AM
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Default Re: Limit of detection

Hi piblanlo,

Do you mean increase your limit of detection in realtime pcr? your requirement is 19/20? with replicate? But you are only getting 17/20?

Is the method validated? and since its process of standardization, does other ppl have the same result or better than you?
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Old 09-30-2011, 08:42 AM
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Default Re: Limit of detection

Hi, thanks for your answer.

I mean decrease the limit of detection. That is, i want to detect a very low number of copies, for example, it would be perfect that my reaction was able to detect 2 copies of my DNA. At this moment, I have achieved a limit of 16 copies/reaction....do you understand me??
the method is not validated, there are some kits of PCR but they detect a number of copies lower than 16....for this reason I need a better result

thanks!!
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  #6  
Old 09-30-2011, 05:49 PM
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Default Re: Limit of detection

hmmm, let just say improve your limit of detection.

let me read up on the smartcycler info will tell you soonest. I need to verify something... my hunch.
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  #7  
Old 10-07-2011, 05:32 AM
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Default Re: Limit of detection

Still checking, BTW if anybody have solution please don't be shy
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