Hi,
after having spent a short while in Indonesia on a project involved in
Jembrana disease of cattle (a retrovirus), I am back in the land of internet
connections. A PhD student in my lab, Loryn Sellner, is trying to develop a
PCR system for detection of alphaviruses in mosquitoes. We are quite a long
way down the track in that we have effectively a one tube, nested RT-PCR for
one such virus. The sensitivity is at the handful of RNA molecules level.
Unfortunately, processing more than 20 mosquitoes in a 1 ml buffer leads to
inhibition, despite the guanidium or RNAzol B extraction of the mosquito
grind. We would like to get to the 1000 mosquitoes/reasonable volume stage
(possibly at the level of 200 mosquitoes/ml). Does anyone know of a
non-mechanical way of 'digesting' the mosquitoes so that their 'guts' become
liberated ? Any thoughts on this matter would be appreciated.
Finally, I am sorry I can't be at ICV, I hope those of you who go get a chance
to link up !! And please make some noise about INTERNET, in particular of
course about bionet.virology
Kindest regards,
Robert
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Dr R.J. Coelen phone 61 - 9 - 389 3915
Dept of Microbiology fax 61 - 9 - 389 2912
University of Western Australia snail Nedlands, 6009
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