I am performing a Lamp PCR protocole using Hydroxy Naphthol Blue, I am using 200uM, I found that the Mg2+ suitable for this protocolo is 6mM, however, every time that I add the dntps in the solution the color purple given by the concentration of Mg2+ starts to change to dark blue, I know that it the dntps in contact with Mg2+ generates a reaction, but how some methodologies use the same reagent and for the control negative the color purple is present, I hope some one could give me some suggestions, my buffer is composed of (NH4)2SO4 10mM, KCl 50mM, Tween-20 0.1%, Tris-HCL 20mM pH 8.8.
I am no sure that this is a problem. Hydroxy naphthol blue is an organic acd that changed colour depending on the absence or presence of metal ions so it is probably normal to get a colour change on adding Mg and the effect may be influenced also by the dntps taking some Mg out of the reaction mix but this is only a problem if it interferes with the reaction
Paul Rutland Thank you so much for this answer, I appreciate it. I will try to perform a better concentration of Mg2+, maybe I should continue with the standardization.
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