Translation by dialysis
This dialysis method is used to increase the yield compared to simple batch method or for proteins that are difficult to synthesize by simple batch.
Yield tends to increase by a factor of about 1.5 with simple batch that is larger than one reaction (110 ul) in our standard protocol , 550 ul for 5 reactions. If the sample yield is smaller than expected with 110 ul of sample, please consider using a sample size of about 550 ul, too.
A general dialysis method can be used in which a translation solution is placed inside the dialysis cup (sample solution) and an amino acid mix is used as an external solution for dialysis. Usually, the volume of dialysis solution is 20 to 100 times larger than inside.
- Dispense 4x diluted Amino Acid Mix (AAMX004) 2ml into floater. AAMX004 is for dialysis substrate exchange layers.
- Set the dialysis cup.
- Dispense 110 μl of the solution prepared by normal translation into the dialysis cup.
- Set the water level of the inner and outer solution to the same level
- Incubate at 16 °C for 10 hours or more ( to 20 hours)
Slide-A-Lyzer dialysis cup (0.1 ml, MWC 3.5K) will fit 24 well titer plate with 9-9.5mm hole. you can make a hole on cap with 9.0 mm drill to fit the dialysis cup and as floater like follows.