Construction of pGGDestTol2LC-sgRNA vectors
Ligate the U6x:sgRNA cassettes into a pGGDestTol2LC vector by mixing the following components: 
Incubate mixture at 37 °C for 20 min. 
Incubate at 16 °C for 15min. 
Incubate at 80 °C for 15min. 
The reaction is ready for transformation (use 5 µl of the ligation and plate 50% of the transformants).
Transform and spread onto ampicillin (100 µg/ml) plates.
