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Team:Illinois/Antibody Receptor Tyrosine Kinase Fusion Notebook
From 2008.igem.org
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July 1, 2008
Made Yeast Media
YPD Medium, per liter:
10g yeast extract
20g peptone
20g dextrose
20g agar(only for plates)
1. Dextrose filter sterilized, the rest autoclaved
2. Weigh nutrients into flask double the volume you want to make, and stir to dissolve
3. Dextrose added to autoclaved media to equivalent of 20 g/L
4. Liquid media placed on bench, plate media placed in 65 degree water bath approximately 5 minutes
5. Poured into plates and allowed to solidify
July 17, 2008
E.Coli Media
LB medium, per liter
10g tryptone
5g yeast extract
5g NaCl
1 mL 1N NaOH
15g (agar for plates)
1. Antibody DNA resuspended in TE buffer, 0.1 μg/μL
2. 5mL LB inoculated with single colony DH5a pro
3. Incubated at 37 degrees overnight
July 18, 2008
Competent cells:
1. 3mL overnight culture of DH5a pro
2. Inoculated into 35mL of LB
3. OD600 checked, want 0.2-0.3
4. Place culture on ice for 3 minutes
5. Spin at 10,000 rpm for 7 minutes, discard supernatant
6. Resuspend in 10 mL cold 30 mM CaCl2, incubated on ice overnight
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