Team:Macquarie Australia/Protocols/ResultsofGibsonsofar110912

From 2012.igem.org

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* T7 Agrobacterium: no colonies were present on the chloramphenicol plates.  
* T7 Agrobacterium: no colonies were present on the chloramphenicol plates.  
* Agrobacterium: no colonies were present on either Kanamycin or Ampicillin plates.
* Agrobacterium: no colonies were present on either Kanamycin or Ampicillin plates.
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 +
{| border="3" cellpadding="4" cellspacing="0" align="center" style="width: 100%; height: 400px"
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|-
 +
! scope="col" colspan="1" style="background-color: gray;"|
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! scope="col"| #1C
 +
! scope="col"| #2K
 +
! scope="col"| #2A
 +
! scope="col"| #3K
 +
! scope="col"| #3A
 +
! scope="col"| #4C
 +
! scope="col"| #5K
 +
! scope="col"| #5A
 +
 +
|-
 +
! scope="row" rowspan="5" width="20%" | 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
 +
 +
|-
 +
! scope="row"| 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
 +
 +
|-
 +
! scope="row"| 3. Addition of Gibson Master Mix (µl)
 +
| 10
 +
| 10
 +
| 10
 +
| 12.9
 +
| 12.8
 +
| 12.7
 +
| 12.9
 +
| 12.8
 +
 +
|-
 +
! scope="row"| 4. Addition of deionised H2O
 +
| 3.3µl
 +
| 3.1µl
 +
| 3.2µl
 +
| nil
 +
| nil
 +
| nil
 +
| nil
 +
| nil
 +
 +
|-
 +
! scope="row"| Outcome
 +
|
 +
|
 +
|
 +
|
 +
|
 +
|
 +
|
 +
|
 +
|}
 +
<br/>

Revision as of 04:11, 11 September 2012




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


Retrieved from "http://2012.igem.org/Team:Macquarie_Australia/Protocols/ResultsofGibsonsofar110912"