Limit of detection

[piblanlo]

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

[piblanlo]

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

[piblanlo]

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

[butters]

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?

[piblanlo]

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!!

[butters]

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.

[butters]

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