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.
