Home FAQs How much linear DNA template and NEBExpress® GamS Nuclease Inhibitor should I add to my reaction?

FAQ: How much linear DNA template and NEBExpress® GamS Nuclease Inhibitor should I add to my reaction?

As a starting point, we suggest adding 250 ng of linear DNA and 1.5 μg GamS (1μL) in a 50 μL reaction to enhance protein yield. The size of the gene and the DNA construct can contribute to the stability of the linear DNA and the rate of transcription. Therefore, different protein targets may require further optimization to determine the best ratio of template and GamS.  
During the experimental setup, it is recommended to add the linear DNA template in the last step to allow GamS to bind and inhibit RecBCD before RecBCD has a chance to act on the DNA.  In addition, the protein synthesis reaction can be scaled to larger volumes, in which case the amount of linear DNA and GamS should also be scaled proportional to the reaction volume.