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 |