Particle bombardment II

Peter Ellis pjie2 at cam.ac.uk
Sun Nov 20 06:19:11 EST 2005


sharp.tool at bigpond.net.au wrote:
>Below Taken from the bio-rad website:
>
>Sample Preparation
>
>The procedure described here prepares sufficient microcarriers for 60 bombardments using 500 micrograms per bombardment. 
>Weigh 30 milligrams of microparticles, either gold or tungsten, into a 1.5ml microfuge tube.
>Add 1 ml 70% ethanol.
>Vortex for 3-5 minutes.
>Allow the particles to soak in 70% ethanol for 15 minutes.
>Pellet the microparticles by briefly spinning in a microcentrifuge.
>Remove and discard the supernatant.
>Wash three times in sterile water, pelleting the microparticles by briefly spinning and discard the supernatant.
>Add 500 microlitres sterile 50% glycerol to bring the concentration to 60 milligrams per ml.
>These microparticles can be stored for up to two weeks, tungsten at -20 0C and gold at -40C or room temperature.
>
>I thought it was strange that it does not include mixing the particles with DNA anyway in the preparation.

Erm.  That's the procedure for preparing the particles for DNA 
coupling.  After this, you then attach the DNA, and finally do the 
bombardment.

>
>Sharp Tool

Are you sure?

Peter


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