Protocol Online logo
Top : New Forum Archives (2009-): : PCR, RT-PCR and Real-Time PCR

Saliva and GCF DNA purification - (Jun/02/2012 )

Dear All,

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)?

-Star 14-

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.

-AquaPlasmid-