I'm using DNA extraction kit (Invitrogen) for isolation of DNA from actinomycetes, it was good for most of the samples, but someof the isolates could not come even i've tried for several times.
These isolates are very hard in nature in the plates. During isolation the cells couldn't be lysed when kept in water bath. So, anyone please suggest me some protocol or ideas how i can get the DNA of these isolates....
Thank you
Hi Zothan,
You may try to grind the biomass of your actinomycetes in liquid nitrogen and use the powder for DNA extraction. Alternatively, crush the biomass in the lysis buffer with a glass rod before starting extraction.
Hope it works!
Tuan Tran
Zothan,
Without knowing the exact kit you used it is hard to know what could be preventing successful DNA extraction. Protocols/ kits that use a combination of mechanical disruption (bead beating on a vortex mixer) with enzymes (proteinase K or similar heated to 55degC) are generally recommended for tough to lyse bacteria and there are lots of these kits about from various companies.
Hope you find a solution
Alison
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