Hi all,

Last week I ordered an oligonucleotide online which I want to use for my CARD-FISH experiment. Yesterday I received the oligo certificate online describing the concentration of the probe delivered and the molecular weight.

Now I would like to dissolve the probe but I am not sure in what volume I should dissolve it. There are online tools to calculate the volume based on the concentration in nmol and the molecular weight of the probe (in g/mol) depending on the desired final concentration. However, the probe is conjugated to a Horse Radish Peroxidase and that is not taken into account in the molecular weight.

So, if I fill in all information needed in the calculator it tells me to dissolve the probe (5.9 nmol / 32.4 µg / OD 1.1) in 648 µL sterile water for a 50ng/µL final concentration. This calculation is based on the molecular weight of the probe which is 5492 g/mol, but does not incorporate the weight of the HRP conjugate at the 5'-end.

When just going out from the 32.4 µg and wanting a 50ng/µL final concentration it seems easy to calculate and you also end up with 648 µL of dissolvent needed. But, since it is the first time and I want to be sure that this is the correct volume I wanted to ask this here.

Thanks!