How should I prepare my linear template DNA and what are your design recommendations for linear DNA templates?

PCR products can be directly used as DNA template for cell-free expression so long as the final glycerol concentration in the reaction is 0.1% or lower and the PCR reactions on average are making enough linear DNA to facilitate the protein yield needed. If a known fixed template concentration is preferred, PCR products should be subjected to a standard PCR clean up procedure with a final elution in molecular biology grade, nuclease-free water. This might also slightly boost the protein yield per reaction. Linear DNA can also be ordered and used directly in myTXTL from several commercial DNA suppliers. Please refer to the applicable myTXTL manual and our Application Note with IDT for information about linear DNA template design requirements, or reach out to techsupport@arbor.daicel.com with questions about specific sequence recommendations.