I use these Universal Eubacteria16S gene Primers Fwd 5’-GGTGAATACGTTCCCGG-3’; Rev 5’-TACGGCTACCTTGTTACGACTT-3’ quite often.

with - iQ™ SYBR® Green Supermix (Bio Rad, Catalog number 170-8880).

Adjust the cycling conditions as follows, 1 cycle: at 95°C for initial denaturation and enzyme activation for 3 min; 40 cycles: at 95°C for denaturing for 15 sec, for annealing at 58°C for 30 sec, for extension at 72°C for 30 sec at; finalize at 4°C.

good luck!

Hi,

in case this is interesting for you I am currently looking for scientist which want to test such a solution which I am developing right now. The solution will have some advantages like absence of bacterial background. Let me know if this is interesting for you

Best Sven

There maybe more than one copy of 16s rDNA.

Delin Liang This is correct and therefore qPCR based quantification is rather a tool to quantify the amount of amplifyable 16S copies. However, for most sample types the predominant strains are known and you can check the number of 16S copies/ Genome. With this in mind you can interpolate the number of genomes/µl and calculate back on your sample input

Sven

Ozkan Gelincik Thank you for the advice, do you have a citation for these primers?

Sven Reister Thank you for the explanation.