Particle bombardment II

Sharp Tool sharp.tool at bigpond.net.au
Sun Nov 20 05:54:52 EST 2005


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" <oliver.berkowitz at anu.edu.au> wrote in message news:437ff781$1 at clarion.carno.net.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

  http://www.rsbs.anu.edu.au/ 

    "Sharp Tool" <sharp.tool at bigpond.net.au> wrote in message news:VLgff.20405$Hj2.18670 at news-server.bigpond.net.au...
    Hi,

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

    Cheers
    Sharp
-------------- next part --------------
An HTML attachment was scrubbed...
URL: http://www.bio.net/bionet/mm/methods/attachments/20051120/4b8ebfd0/attachment.html


More information about the Methods mailing list