Table 10.

Components of Golden Gate reaction and thermal conditions.

Item	Concentration	Amount	Temperature	Time	Cycle
Inserts *		(100 ng)	37 °C	5 min	40
50X oligonucleotide **	0.5 μM	0.3 μL	20 °C	10 min
10X T4 Ligase Buffer		1.5 μL	20 °C	1 h	1
AarI	2 U/μL	0.5 μL	65 °C	20 min	1
T4 Ligase	350 U/μL	1 μL	8 °C	Hold	1
Distilled Water		to 15 μL

* Every insert. ** Must be added owing to efficiency.