Team:Macquarie Australia/Protocols/GibsonAssembly

From 2012.igem.org

(Difference between revisions)
Line 41: Line 41:
|-
|-
! scope="row"|  
! scope="row"|  
-
|
+
| nil
-
|
+
| nil
| 2µl Deino_C
| 2µl Deino_C
| 2µl Deino_C
| 2µl Deino_C
Line 52: Line 52:
|-
|-
! scope="row"|  
! scope="row"|  
-
|  
+
| nil
-
|
+
| nil
| 2µl Deino_D
| 2µl Deino_D
| 2µl Deino_D
| 2µl Deino_D
Line 63: Line 63:
|-
|-
! scope="row"|  
! scope="row"|  
-
|  
+
| nil
-
|
+
| nil
| 2µl Deino_E
| 2µl Deino_E
| 2µl Deino_E
| 2µl Deino_E
Line 98: Line 98:
| 3.1µl
| 3.1µl
| 3.2µl
| 3.2µl
-
|
+
| nil
-
|
+
| nil
-
|
+
| nil
-
|
+
| nil
-
|
+
| nil
|}
|}

Revision as of 10:11, 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 2µl Deino_C 2µl Deino_C 2µl Agro_C 2µl Agro_C 2µl Agro_C


nil nil 2µl Deino_D 2µl Deino_D 2µl Agro_D 2µl Agro_D 2µl Agro_D


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



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