"
Team:Macquarie Australia/Protocols/GibsonAssembly
From 2012.igem.org
(Difference between revisions)
| Line 6: | Line 6: | ||
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. | ||
| - | {| border=" | + | {| border="5" cellpadding="0" cellspacing="0" align="center" |
|- | |- | ||
! scope="col"| Instruction | ! scope="col"| Instruction | ||
| Line 28: | Line 28: | ||
| 2µl Agro_A | | 2µl Agro_A | ||
| - | |||
! scope="row"| 2µl Hemo_B | ! scope="row"| 2µl Hemo_B | ||
| + | |- | ||
| 2µl Hemo_B | | 2µl Hemo_B | ||
| 2µl Hemo_B | | 2µl Hemo_B | ||
Revision as of 10:08, 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
| |||
| 2µl Deino_C | 2µl Deino_C | 2µl Agro_C | 2µl Agro_C | 2µl Agro_C
| |||||
| 2µl Deino_D | 2µl Deino_D | 2µl Agro_D | 2µl Agro_D | 2µl Agro_D
| |||||
| 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 |
