TX-TL Reaction Mix Example

The following is an example TX-TL reaction following the standard used by the Murray Lab at Caltech. In general, the salt concentrations for each batch of TX-TL are calibrated separately. Additionally, it is worth noting that the final salt concentration of the entire mixture is difficult to calculate due to the complicated set of salts in the energy buffer which isn't described here.

Total Reaction Volumes are typically 5-20uL (with 10uL being the suggestion for use in a 384 well plate).

Ingredient	% (by Volume)
Cell Extract	33%
Amino Acid Mix	12.5%
Energy Buffer Mix	7.5 %
PEG-8000	5%
Remaining (for Salt and DNA)	42%

Salt, DNA, Water and GamS are all added to reactions in variable amounts. Standard ranges are given here; the concentrations listed are the final concentration added to the reaction volume.

Ingredient	Final Concentration Added
Magnesium-Glutamate	2-10 mM
Potassium-Glutamate	50-150 mM
DNA Constructs	.5 - 20 nM
Gam S (to protect Linear DNA)	2-4 uM
Water	The remaining volume