Saliva and GCF DNA purification - (Jun/02/2012 )
I would like to know what is the best DNA purification kit that you recommend for isolating different strains of oral bacteria from either saliva and gingival cervicular fluid (GCF)?
It works for bacterial and buccal DNA extraction, but I am not 100% sure if AquaGenomic would work for DNA extraction from the oral bacterial strains you are after. For saliva DNA extraction, you would add some proteinase k to the AquaGenomic solution, mix it with the saliva, incubate at 60*C for 1hr and then at 95*C 15min to kill the proteinase K, spin to get the clear lysate, and then precipitate the DNA with isopropanol. If this does work, you could try adding a lysozyme digest step prior to the AquaGenomic/proteinase k step to break down the bacterial cell walls first (be sure to use TE buffer to keep the saliva pH ~8-9 for optimal lysozyme activity). If you are interested, we would be happy to send you a trial sample of AquaGenomic.