Insert foreign GST tag protein sequence in the protein needs a checklist from Primary steps of DNA Engineering to make sure about the latest result in protein. These are important key for high quality GST tagging process.

Insert DNA must possess an open reading frame and should be less than 2 kb long to make sure about size, weight and the latest result. Wehether subcloned from another vector or amplified by PCR, the insert must have ends that are compatible with the linearized vector ends. Using two different restriction enzymes will allow for directional cloning of the insert into the vector. This process will help engineer for latest processes and protocols running on the DNA sequence. Directional cloning will optimize for inserts in the correct orientation.

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