Lab Manual
biological reagents
Cell Free Systems
In Vivo Systems
misc methods
CELL FREE SYSTEMS > General Cell Free Translation  

> Reticulocyte Lysate and Wheat Germ Extracts

  > tRNA Crosslinking > Preparative Aminoacylation of Pure tRNA

> Phenol Extraction of tRNA

    > Generation of Integration Intermediates

> Immunoprecipitation of translation products


> Translation in RRL


> History and evolution of cell-free protein synthesizing systems


> Import Into Mitochondria


> Wheat Germ Cell-Free Translation


> E coli Translation




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  1. Assemble mm. Add lysate tRNA in the dark. Wrap tubes in foil. Incubate 24oC, 60 minutes.
  2. Add equal volume (100ml) 1X retic and place into cuvette. (doesn't have to be UV grade).
  3. Flash for 15 sec. at 300-400nm band width (double filters) (photolysis machine in Gerber's lab has 1000W bulb).
  4. Pass samples over CL 2B column (2 ml bed volume) and collect void volume (This will separate targetted molecules from other junk) (High salt will remove rb x-links) for 2 ml columns void volume ~ 700--> 1000ml (drop #'s will depend on drop size) . 55 (drop size---> drops 13-19 (~400ml ; 7 drops)).