About scale-up
In this protocol, 1 translation reaction is 110 ul, but it can be scaled up. In that case, please increase the reagents for translation reaction at the same ratio.
e.g.) when scaling up 110 ul of translation reaction to 220 ul of 1 reaction is
e.g.) when scaling up 110 ul of translation reaction to 220 ul of 1 reaction is
Wheat germ extract 10 ul >> 20 ul
Amino acid mix 20 ul >> 40 ul
mRNA 80 ul >> 160 ul
Note that the target protein yield may be improved by scaling up.
e.g.) From a translation reaction of 110 ul with a yield of 10 ug to a translation reaction of 550 ul, the target protein yield is approximately 70 ug and the yield per volume of the reaction solution is improved.
Therefore, if you want to improve the protein yield after adjusting the amount of mRNA per reaction in the standard protocol, please try in the following order.
- Increase translation reaction time from 10 to 20 hours
- Scale-up
- Dialysis method