FAQ: The protocol recommends shaking the reactions vigorously. Is this necessary?

It is not necessary to shake reactions vigorously if there is sufficient headspace in the tubes containing the reactions. Shaking may improve yield, but it is not required. If you choose to shake your reactions, we recommend shaking at 800-1000 rpm in a temperature-controlled mixer (e.g. Eppendorf ThermoMixer or equivalent).