Golden Gate Assembly

Updated 5/25/2018 02:02 PM

by Boston University iGEM WL 2018 (created by Linda Luo)

Introduction

Golden Gate Assembly as taught by Meghan!

Make sure all plasmids you need have their concentrations labelled on their tubes (If not, get their concentrations on the nanodrop)

Table2

For the water and DNA mix, first see how much DNA you need

For pYTK's, you need 150ng/trans

For pKL0's, you need 150ng/trans

For pKLE's, you need 20ng/trans

So, use the concentration of the plasmid to determine what volume of the plasmid you need (does not have to be exact)

Put the prepared mix in a thermocycler (Golden Gate 1 Program) for 3 min at 37 degrees and 3 for 16 degrees 20 cycles