Team:Macquarie Australia/Protocols/GibsonAssembly

From 2012.igem.org

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Revision as of 09:51, 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