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Team:Cornell/Week 9
From 2011.igem.org
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July 31st - August 6th
Sunday
Monday
Tuesday
Wednesday
Thursday, August 4
Afternoon lab work done by: Claire Paduano, Charlie Chung
- Objective
- Ligation of GFP/RFP/VioE + avitag + backbone
- Ligation
- GFP + Avi-Tag + Backbone
- 10.6μL pZE12 +avitag vector backbone
- 3.6μL GFP
- 2.9μL H2O
- 2.0μL T4 DNA Ligase Buffer
- 1.0μL T4 DNA Ligase
- 20μL Total
- RFP + Avi-Tag + Backbone
- 10.3μL pZE12 + avitag vector backbone
- 6.7μL RFP
- 2.0μL T4 DNA Ligase Buffer
- 1.0μL T4 DNA Ligase
- 20μL Total
- VioE + Avi-Tag + Backbone
- 2.3μL H2O
- 4.1μL VioE
- 10.6μL pZE12 + avitag vector backbone
- 2.0μL T4 DNA Ligase Buffer
- 1.0μL T4 DNA Ligase
- 20μL Total
- In all above reactions, volume of plasmid vector added corresponds with 100ng backbone.
- Prepare control ligation reactions (inserts replaced with H2O) for each of the above constructs.
- Same volume of vector backbone, buffer, ligase
- Volumes of all inserts (i.e. gene or primer pairs) go toward volume of H2O
- After ligation setup, incubate in 16°C waterbath overnight.
