Extraction of DNA from termite gut flagellates (View forum version)


Posted 13 June 2013 - 04:52 AM

How to extract DNA efficiently from termite gut protozoans?? I attempted several times by following different protocols but got no results..:(


Posted 13 June 2013 - 08:23 AM

Which protocols did you tried out? And do you need pure protozoan DNA or a mixture with termite DNA is okay (e.g. if you have specific primers for the protozoan DNA so that the other is no problem)?
I guess for the former protocols are available (perhaps dissecting termites and removing gut and then flushing out protozoan content with a buffer (never did this, so no real idea)....), and for the latter a normal DNA extraction is surely fitting and many protocols exist (e.g. salting out DNA, Phenol-Chloroform, kits,...).


Posted 30 July 2013 - 01:42 AM

Sir! I have tried all these conventional methods you have mentined but got no results. Now I am using Master Pure Complete DNA and RNA purification kit. I want to collect DNA from the flagellated specific protozoans P. grassii. I tried this protocol to extract DNA. But now when run that sample on agarose gel i was shocked, i got no results even the ladder i used was not visible.


Posted 30 July 2013 - 05:24 AM

This suggests that you have a problem with running your gel, not with the extraction. I would practice making and running gels with the ladder only until it works. We could help if you provide us with details as to how you are doing it.


Posted 30 July 2013 - 09:01 AM

As the genome size of these flagellates is very less thats why i am running it on 2 % agarose gel.


Posted 30 July 2013 - 10:41 AM

The genome size of any chromosome in any organism is much larger than can be resolved on an agarose gel.


Posted 30 July 2013 - 12:10 PM

So then what do you suggest??


Posted 30 July 2013 - 12:29 PM

I'd suggest that you describe what your real goals and plans are, and we can perhaps help you design experiments to achieve them.