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Particle bombardment II

Particle bombardment II - Protocols and Methods Forum

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  #1  
Old 11-18-2005, 09:01 AM
Sharp Tool
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Default Particle bombardment II



Hi,

What is the role of spermidine and calcium chloride in preparing DNA-coated particles for bombardment?

Cheers
Sharp

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  #2  
Old 11-20-2005, 04:21 AM
Oliver Berkowitz
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Default Particle bombardment II

Hi,

protecting the DNA and ppt to the particles. BTW Bio-Rad has a quite nice website for particle bombardment.

cheers
Oliver

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"Sharp Tool" <[Only registered users see links. ].au> wrote in message news:VLgff.20405$[Only registered users see links. ].au...
Hi,

What is the role of spermidine and calcium chloride in preparing DNA-coated particles for bombardment?

Cheers
Sharp
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  #3  
Old 11-20-2005, 10:54 AM
Sharp Tool
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Default Particle bombardment II

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.

Sharp Tool


"Oliver Berkowitz" <[Only registered users see links. ].au> wrote in message news:437ff781$[Only registered users see links. ].au...
Hi,

protecting the DNA and ppt to the particles. BTW Bio-Rad has a quite nice website for particle bombardment.

cheers
Oliver

--
_______________________________

Dr. Oliver Berkowitz
The Australian National University
Research School of Biological Sciences
Environmental Biology Group
GPO Box 475
Canberra ACT 0200
Australia

P: +61 - (0)2 - 61254549
F: +61 - (0)2 - 61254919

[Only registered users see links. ]

"Sharp Tool" <[Only registered users see links. ].au> wrote in message news:VLgff.20405$[Only registered users see links. ].au...
Hi,

What is the role of spermidine and calcium chloride in preparing DNA-coated particles for bombardment?

Cheers
Sharp

Reply With Quote
  #4  
Old 11-20-2005, 11:19 AM
Peter Ellis
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Default Particle bombardment II

[Only registered users see links. ] wrote:

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


Are you sure?

Peter
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