Protocol Online logo
Top : Forum Archives: : Molecular Biology

DNA extraction from soil - (Apr/13/2007 )

I am trying to isolate DNA directly from soil procured from a depth of 40m. After following a protocol which involved lysozyme amd protenase K aided lysis and subsequent DNA purification and precipitation i got UV spectra with peak at 270 nm. This peak can be attributed to the presence of contaminants (probably phenol which i used to extract the aqueous phase during DNA purification). It does not mean i didnot get anything at 260 nm. I got but it was not a peak. Can i deduce that my protocol worked but great deal of optimisation is needed? Can you friends suggest me how to decrease the contamination in the protocol?

-alokmay-

Have no experience of extarction but some people in my previous lab used this protocol, from zhou et al

http://aem.asm.org/cgi/content/abstract/62...pe2=tf_ipsecsha


They also used with success a kit

QUOTE (alokmay @ Apr 13 2007, 03:32 PM)
I am trying to isolate DNA directly from soil procured from a depth of 40m. After following a protocol which involved lysozyme amd protenase K aided lysis and subsequent DNA purification and precipitation i got UV spectra with peak at 270 nm. This peak can be attributed to the presence of contaminants (probably phenol which i used to extract the aqueous phase during DNA purification). It does not mean i didnot get anything at 260 nm. I got but it was not a peak. Can i deduce that my protocol worked but great deal of optimisation is needed? Can you friends suggest me how to decrease the contamination in the protocol?

-tertu-

I also work with DNA from soil. Sorry,I haven't know the answer yet, since I haven't had the pure yield DNA from soil as well. Anyway, what kind of method did you use? is the DNA yield high?I already used Zhou et al and Dong et al, but I found dificulities in obtaining the high yield of DNA and pure DNA as well.

-rif_researcher-