Team:Macquarie Australia/Protocols/ResultsofGibsonsofar110912
From 2012.igem.org
Results
- T7 Heme oxygenase (chloramphenicol): This biobrick assembly was successful. The plasmids will be sequenced and re-transformed into BL21 DE3 Rosetta & Star cells.
- Heme oxygenase (Kanamycin): Incubation is to be performed today and plasmid preparation using the Qiagen protocol will be performed. No colonies were present on the T7 minus heme oxygenase on ampicillin.
- Deinococcus: Plasmid sequencing will be performed and transformation was successful on kanamycin plates. The Deinococcus on the kanamycin was unsuccessful.
- T7 Agrobacterium: no colonies were present on the chloramphenicol plates.
- Agrobacterium: no colonies were present on either Kanamycin or Ampicillin plates.
#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 |
Outcome (11/09/2012) |