Team:Macquarie Australia/Protocols/GibsonAssembly
From 2012.igem.org
(Difference between revisions)
Line 29: | Line 29: | ||
|- | |- | ||
- | | 2µl Hemo_B | + | | 2µl Hemo_B |
- | | 2µl Hemo_B | + | | 2µl Hemo_B |
- | | 2µl Hemo_B | + | | 2µl Hemo_B |
| 2µl Deino_B | | 2µl Deino_B | ||
| 2µl Deino_B | | 2µl Deino_B | ||
- | | 2µl Agro_B | + | | 2µl Agro_B |
- | | 2µl Agro_B | + | | 2µl Agro_B |
- | | 2µl Agro_B | + | | 2µl Agro_B |
Line 45: | Line 45: | ||
| 2µl Deino_C | | 2µl Deino_C | ||
| 2µl Deino_C | | 2µl Deino_C | ||
- | | 2µl Agro_C | + | | 2µl Agro_C |
- | | 2µl Agro_C | + | | 2µl Agro_C |
- | | 2µl Agro_C | + | | 2µl Agro_C |
Line 56: | Line 56: | ||
| 2µl Deino_D | | 2µl Deino_D | ||
| 2µl Deino_D | | 2µl Deino_D | ||
- | | 2µl Agro_D | + | | 2µl Agro_D |
- | | 2µl Agro_D | + | | 2µl Agro_D |
- | | 2µl Agro_D | + | | 2µl Agro_D |
Revision as of 10:15, 5 September 2012
Gibson Assembly Protocol
Before entering the lab, the assigned Gibson Assembly team for the day organised agar plate requirements, concentration calculations and equipment requirements.
Instruction | #1C | #2K | #2A | #3K | #3A | #4C | #5K | #5A |
---|---|---|---|---|---|---|---|---|
1. Addition of 20ng Gene Block Fragments in appropriate tube | 2µl Hemo_T7A | 2µl Hemo_A | 2µl Hemo_A | 2µl Deino_A | 2µl Deino_A | 2µl Agro_T7A | 2µl Agro_A | 2µl Agro_A |
2µl Hemo_B | 2µl Hemo_B | 2µl Hemo_B | 2µl Deino_B | 2µl Deino_B | 2µl Agro_B | 2µl Agro_B | 2µl Agro_B
| |
nil | nil | nil | 2µl Deino_C | 2µl Deino_C | 2µl Agro_C | 2µl Agro_C | 2µl Agro_C
| |
nil | nil | nil | 2µl Deino_D | 2µl Deino_D | 2µl Agro_D | 2µl Agro_D | 2µl Agro_D
| |
nil | nil | nil | 2µl Deino_E | 2µl Deino_E | 2µl Agro_E | 2µl Agro_E | 2µl Agro_E | |
2. Addition of 0.05 pmol of vector | 2.7µl PSB-1C3 | 2.9µl PSB-1K3 | 2.8µl PSB-1A3 | 2.9 µl PSB-1K3 | 2.8 µl PSB-1A3 | 2.7µl PSB-1C3 | 2.9µl PSB-1K3 | 2.8µl PSB-1A3 |
3. Addition of Gibson Master Mix (µl) | 10 | 10 | 10 | 12.9 | 12.8 | 12.7 | 12.9 | 12.8 |
4. Addition of deionised H2O | 3.3µl | 3.1µl | 3.2µl | nil | nil | nil | nil | nil |