FAQ: Why do many of the published Golden Gate Assembly articles feature precloned inserts as opposed to inserts generated by PCR?

Precloned inserts allow stable storage of inserts and make it easy to calculate the molarity of inserts, while using amplicon inserts saves time. Stable storage of amplicon inserts is important. Single insert cloning/assemblies can use unpurified amplicons, all other amplicons should be purified, for example by spin column protocols. For long term storage at –20°C, store DNA in 10 mM Tris (pH 8.5), 0.1 mM EDTA.