Team:Macquarie Australia/Protocols/ResultsofGibsonsofar110912

From 2012.igem.org

Revision as of 04:11, 11 September 2012 by Ehoyer (Talk | contribs)




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