Team:Macquarie Australia/Protocols/GibsonAssembly2

From 2012.igem.org



9 Gibson Assembly Reactions (2nd Attempt)

3A 3K 3C 4A 4K 4C 5A 5K 5C
1. Addition of 20ng gene fragments in appropriate tube 2 μL each of Deino_A, Deino_B, Deino_C, Deino_D, Deino_E 2 μL each of Deino_A, Deino_B, Deino_C, Deino_D, Deino_E 2 μL each of Deino_A, Deino_B, Deino_C, Deino_D, Deino_E 2 μL each of AgroT7_A, Agro_B, Agro_C, Agro_D, Agro_E 2 μL each of AgroT7_A, Agro_B, Agro_C, Agro_D, Agro_E 2 μL each of AgroT7_A, Agro_B, Agro_C, Agro_D, Agro_E 2 μL each of Agro_A, Agro_B, Agro_C, Agro_D, Agro_E 2 μL each of Agro_A, Agro_B, Agro_C, Agro_D, Agro_E 2 μL each of Agro_A, Agro_B, Agro_C, Agro_D, Agro_E
2. Addition of 0.05pmol of vector 2.8 μL PSB-1A3 2.9 μL PSB-1K3 2.7 μL PSB-1C3 2.8 μL PSB-1A3 2.9 μL PSB-1K3 2.7 μL PSB-1C3 2.8 μL PSB-1A3 2.9 μL PSB-1K3 2.7 μL PSB-1C3
3. Addition of Master Mix (μL) 12.8 12.9 12.7 12.8 12.9 12.7 12.8 12.9 12.7