Genetic Immunization Bullet Preparation

2015-01-13

azim58 - Genetic Immunization Bullet Preparation

the protocol of these files can be found on the shared drive
"S:\Research\GeneVac\Protocols\Standard Operating Procedure for Making
Bullets - CaCl2 revised_9-26-2011.doc"
"S:\Research\GeneVac\Protocols\nanogold experiment design-updated
june6,2012.xls"

Protocol

* Take the calculated aliquot of colloid gold and transfer it into a
  fresh plastic tube.
* Add the calculated aliquot of DNA to the colloid gold
* Mix. The color of the solution should change to more violet tone.
  Leave at RT for 5 min. Aggregate particles often start to form
* Add the calculated aliquot of charged micro gold to the mix.
* Mix occasionally at RT for 10min. Gold sedimentation rate should
  increase and supernatant discolorate. Mix at about 3 min intervals
* Briefly spin for 1 min 1000 rpm gold by centrifugation
* Aspirate the supernatant
* Resuspend the gold pellet in the residual liquid and add 1ml of
  ethanol to the slurry while vortexing
* Briefly spin gold by centrifugation
* Aspirate the supernatant
* Briefly spin gold again and aspirate the residual ethanol
* Resuspend the gold pellet in the calculated amount of butanol and
  transfer into Teflon tubing
* Let the gold settle in the tubing for 5-10 min
* Drain the butanol and dry the gold by flowing N or Ar gas gently
  through the tubing
* Cut tubing into bullets with tube cutting device (1/2" pieces)
* Store over dryrite.

Here's an example protocol Felicia and I used on 6-18-12
"F:\kurt\storage\CIM Research Folder\DR\2013\5-27-13\wiki_download\bullet
preparation protocol 2 6-18-12.pdf"

azim58wiki: