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CELL FREE SYSTEMS > TRANSLATION > E coli Translation

> Translation Protocol

     

> Sample Protocol

     

> Troubleshooting

     

> Lysate Preparation of S-30

     

> Membrane-Free Ribosomes (mfrs)

     

> Ribosomal Pellet

     

> Dialysis Tubing Work-up

     

> Membrane-Free Translation System

      > Buffers and Reagents

 

Dialysis Tubing Work-up

  1. In 4L beaker, add 2L of 5% bicarbonate, 1mM EDTA solution (2L solution: 100g of NaHCO3, 0.75 g Na4EDTA).Cut the dialysis tubing into appropriate length and put into the bicarbonate solution.Heat to nearly boiling (boil gently) for half hourRinse with distilled water three times.Gently boil in 2L 0.1% SDS (2g SDS in 2L water or 10mL 20% SDS).Rinse with distilled water six times.Gently boil in 2L of 5% bicarbonate, 1mM EDTA solution for 30 min.Gently boil dd H2O. Rinse with distilled water three times.
  2. Repeat step 8.

Store at 4°C in dd H 2O.. Keep nuclease free.