[Drosophila] Re: Dros Digest, Vol 32, Issue 10

Gurudatta Baraka via dros%40net.bio.net (by jaiganesha from gmail.com)
Fri Dec 28 12:17:14 EST 2007


The larval tissue is sticky if u do not add the detergent -and forms clumps
or stick to poor quality eppendorfs ( silocanised brands are good )
usally we use PTX ( 0.1% triton X 100 ) as blocking solution at 4 degree
storage.
during changes use the tips rinsed once in this solution while transferring
the tissue ( it is better use 1000ul tip or cut tips for p200)
but i do not advice storing the samples in this solution for long time .,
they damage the tissue


another thing is gut is the soruce of lot of bacterial contamination !!! so
need to take care of theat as well !! i hope u can also use the sodium azide
in solutions  ( hezardous to humans too . so caution while handling!!)

another reason could be that the tissue is not properly fixed !! check the
fixative parafomaldehyde  ( which may be old and not working ???)

i hope this would help to sortout the problem

with best reagards
Gurudatta

On Dec 28, 2007 12:03 PM, <dros-request from oat.bio.indiana.edu> wrote:

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>   1. handeling the drosophila gut (gayatri priya)
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> Date: Thu, 27 Dec 2007 09:57:50 +0000 (GMT)
> From: gayatri priya <gayatripriya13 from yahoo.com>
> Subject: [Drosophila] handeling the drosophila gut
> To: dros from magpie.bio.indiana.edu
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> HI. i am working with gut of drosophila. i would like
> to know about the way to handle and store the fixed
> guts. i am storing the fixed guts (approx 10-15 in
> number) in an eppendorf tube with PBS. the problem i
> face is that while i pipette out the sample, it gets
> sheared (as i need the entire gut for my investigation
> purpose). also sugggest a way to section the gut (to
> make a vertical tear in its surface) for antibody
> staining purpose. as i am totally new to the area, i
> would be greatful if you can guide me with the whole
> thing (also as soon as possible),
>
> thank you
> gayatri
>
>
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