2x Isothermal Assembly Mixture
Preparing 2x Isothermal Assembly Aliquots
A manuscript detailing our method, including preparation of the 2x isothermal assembly aliquots, is forthcoming. For now we recommend using the Megason Lab's Protocol with one minor modification:
The protocol recommends combining a 320 uL aliquot of "5x Isotherm Buffer" with various enzymes and 700 uL of ddH2O, then making 15 uL aliquots. We recommend instead adding 300 uL of ddH2O. This yields a 2x assembly mixture, of which we recommend aliquotting 10 uL.