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Team:Macquarie Australia/Protocols/GibsonAssembly
From 2012.igem.org
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Before entering the lab, the assigned Gibson Assembly team for the day organised agar plate requirements, concentration calculations and equipment requirements. | Before entering the lab, the assigned Gibson Assembly team for the day organised agar plate requirements, concentration calculations and equipment requirements. | ||
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! scope="col" colspan="1" | Instruction | ! scope="col" colspan="1" | Instruction | ||
Revision as of 10:18, 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 |
