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"