Lamp loop mediated isothermal amplification question. Lowest concentration of DNA to get positive reaction with dye sybr green or Eva green

Leonardo Pantoja Munoz @Leonardo-Pantoja-Munoz

15 May 2017 1 5K Report

I am doing loop mediated isothermal amplification.

I need to know what is the lowest concentration of DNA that gives positive fluorescence using sybr green or Eva green, both specific for sDNA

This because my master mix gives false positives when only adding primers and target DNA, but before incubation/amplification

Leonardo Pantoja Munoz

Hi Mahbubul

Thanks for your replies

I have tried that and I know for sure there is not contamination

I have done this set of samples,

only RNAse free water + dye
RNAse free water + dye +primers and target DNA
RNAse free water + all lamp reagents + primers and target DNA

All samples before lamp incubation done at 4C degrees

only RNAse water gives negative results (no fluorescence)

the other 2 samples give positive fluorescence

As you mentioned I need to optimise the concentration of my primers

Thanks

Leonardo

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