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Extraction of DNA from termite gut flagellates

Protozoan flagellates DNA Extraction Termites

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7 replies to this topic

#1 hamidicup

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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..:(

#2 hobglobin

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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,...).
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#3 hamidicup

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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.



#4 phage434

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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.



#5 hamidicup

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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.



#6 phage434

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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.



#7 hamidicup

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Posted 30 July 2013 - 12:10 PM

So then what do you suggest??



#8 phage434

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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.







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