I am using the QIAGEN kit for DNA isolation from stool for bacteria detection. I am also using Zirconia beads for mechanical destruction of the bacterial cells as a step in DNA isolation. I don't have a bead beater , Omnilyser or similar instruments. Would vortexing the stool sample containing the beads be enough? If yes, then for how long do you suggest?
bead-beater is strongly recommended.
however MoBio company says in kit manual to "secure tubes horizontally on a flat-bed vortex pad with tape. Vortex at maximum speed for 10 minutes"
check link
they also sell adapter. it seems that the tubes must be placed horizontally to allow the longest amplitude for beads movement.
http://www.mobio.com/fecal-dna-isolation/powerfecal-dna-isolation-kit.html
Yes, absolutely. The BD method involves vortexing for 5 minutes at full speed. The only issue might be the size of the beads that you use. Have a look at their patent; I think that it says that there is a mix of bead sizes between 100 nm and 1.2 um. However, others have found comparable results using other bead sizes. Try a PubMed search.
Hi
Some high speeds vortexes also can be used for mechanical disruption. They looks similar to conventional vortex, however they work at high rpm
One good example is
http://www.scientificindustries.com/disruptorgenie.html
best regards
Thank you all for your answers, suggestions and advices.
Mr. Nick.....The beads i am using are a mixture of 0.1 mm and 0.5mm .
I always use a bead beater, with or without beads the disruption seems to produce good quality DNA yield and purity. You could also try a lysozyme pre-treatment step if you do not have access to a bead beater as I have read that that also works quite well
Vortex can't replace a bead beater
Article Use of Accuracy Profile Procedure to Validate a Real-Time PC...
vortexers work great, but you typically need 10 min for efficient sample disruption. so dont do it by hand of course, use the adapter to plug in several bead tubes- and shake them. not the vertical one, but horizontal one- its crucial.
Kido, H., M. Micic, D. Smith, J. Zoval, J. Norton, and M. Madou. 2007. A
novel, compact disk-like centrifugal microfluidics system for cell lysis and
sample homogenization. Colloids Surf. B Biointerfaces 58:44–51.
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