You need to determine the concentration of your DNA first. You can use a nanodrop, Qubit system or Agilent Bioanalyzer. I prefer the Qubit as it is quite accurate. The nanodrop is quicker but it can be affected by contamination.

So for instance, if your concentration is 50ng/uL, then you would need 1uL for your PCR reaction as 1uL of DNA contains 50ng.

Hi Syafiqah Mhd Jamal

Usually we dissolve DNA into suitable buffer for PCR work. After dissolving in buffer, then we quantify the DNA in ng or ug by Nanodrop spectrophotometer. Actually nanodrop gives us the concentration of DNA/RNA per ul of solution. As for example if your nanodrop gives you 100ng/ul of DNA/RNA solution; it means that, your 100ng of DNA is present/ul of your solution; then you can calculate the total yield of your DNA/RNA from your volume.

Thanks

Hi Syafiqah Mhd Jamal

First, you have to know your DNA template concentration. If you have qubit, or nanogram, or another analyzer, that will be good. The result of this analyzer will be on ng or ug per volume (uL).

If your result is 50ng/uL of DNA template, it means you need to add 1 uL of DNA template to the 49uL of reaction to obtain 50uL PCR reaction.

I usually use this in calculation.

uL of the template = (ng) DNA template I need/ (ng) DNA template result by qubit.

= 50/50

= 1uL

Thank you everyone for the answers! It's helpful!

Yes, depending on your DNA concentration (which have to be determined) you can calculate the volume (µl) of amount (ng) you have to add to your PCR mix.