Request for protocols for storage of ticks for DNA isolation

Paul Neese thetick at nando.net
Sat Oct 21 23:39:43 EST 1995


Mr M.R. Owen wrote:
> 
> I am planning on collecting ticks from Africa over the next 
year.  The ticks
> will be stored at room temperature for about 8 months, after 
which I would
> like to look for Babesia DNA in the ticks by PCR.  Has anyone 
any good ideas
> for the best way to preserve the ticks to maximise my chances 
of recovering
> good quality DNA when the samples get shipped over to me  in 
the UK?
> 
> I will also be collecting bovine blood and brain samples.  
Any ideas on the
> best preservation techniques for these too would be grately 
appreciated.
> 
> Thanks in anticipation
> Luis Neves
> email lneves at liverpool.ac.uk

When I was working on my Ph.D. at Oklahoma State University, a 
graduate student in oour lab was working on a project to 
develop a method for identifying greenbug and blue alfalfa 
aphid biotypes using RAPD-PCR.  Part of the study involved 
determining the best method for storing specimens for later 
PCR.  She found that the same primer (10-mers in this case) 
would yield different banding patters corresponding to 
different storage conditions, and that the most harsh storage 
condition (i.e. room temp) yielded the least amount of 
DNA(faint bands when visualized with ethidium bromide)and a low 
number of bands.  This was tested with over 60 10-mers.  I 
would suggest that freezing the specimens would be the best 
form of storage, preferably at -80 degrees C, then -20 degrees, 
and so on.  Storage in EtOH yielded results somewhere between 
room temp storage and freezing.  However, as I mentioned, using 
the same primer under different conditions yielded different 
banding pattern. Thus, comparision of bands between samples 
stored under different conditions using the same primer are 
essentially useless.  I hope this helps.



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