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Team:Yale/Notebook/Week4
From 2011.igem.org
(→Notebook: Week 4) |
(→Notebook: Week 4) |
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''Monday'' | ''Monday'' | ||
| + | |||
| + | - Made glycerol stock of Canada cells. | ||
- Made 1 L LB, completed 1/40 dilution of ZeAFP (added Amp), shook for 3 hrs, checked OD until .78 OD reached | - Made 1 L LB, completed 1/40 dilution of ZeAFP (added Amp), shook for 3 hrs, checked OD until .78 OD reached | ||
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- Divided 1 L into 6 separate 100 mL solutions, half with IPTG, induced for 5 hours | - Divided 1 L into 6 separate 100 mL solutions, half with IPTG, induced for 5 hours | ||
| - | - Centrifuged 4 100 mL solutions, saved supernatant/pellet for 2; for the other 2, we completed the following protocol: | + | - Centrifuged 4 100 mL solutions, saved supernatant/pellet for 2 (frozen at -20); for the other 2, we completed the following protocol: |
1) Washed with 50 mL water | 1) Washed with 50 mL water | ||
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2) Resuspend in 100 mL ice-cold water | 2) Resuspend in 100 mL ice-cold water | ||
| - | 3) | + | 3) Divided 1ml aliquots into 8 eppendorf tubes (half induced half uninduced) |
| + | |||
| + | 4) Added 100 and 10ul to IPTG and non IPTG plates (4 total) and grew at room temperature. Froze eppendorft tubes. Tomorrow we will unfreeze them for 6 hours on ice, then 1 hour at 4C, then replate to compare colony growth - essentially a survival assay after frozen : before frozen number of viable cells (this is obviously just a test assay) | ||
- Plated 6 samples of the remaining 2 100 mL solutions, using 10 and 100 microliters on IPTG, LB, Amp plates and non-IPTG, LB, Amp plates and stored 1 of each concentration in 0 degrees, 4 degrees, and room temperature | - Plated 6 samples of the remaining 2 100 mL solutions, using 10 and 100 microliters on IPTG, LB, Amp plates and non-IPTG, LB, Amp plates and stored 1 of each concentration in 0 degrees, 4 degrees, and room temperature | ||
| - | |||
''Tuesday'' | ''Tuesday'' | ||
- Finished and presented project defense presentation to Sackler Institute and our advisers. | - Finished and presented project defense presentation to Sackler Institute and our advisers. | ||
| - | + | - Obtained TCA protein precipitation protocol. | |
''Wednesday'' | ''Wednesday'' | ||
Revision as of 20:48, 7 June 2011
Notebook: Week 4
Monday
- Made glycerol stock of Canada cells.
- Made 1 L LB, completed 1/40 dilution of ZeAFP (added Amp), shook for 3 hrs, checked OD until .78 OD reached
- Divided 1 L into 6 separate 100 mL solutions, half with IPTG, induced for 5 hours
- Centrifuged 4 100 mL solutions, saved supernatant/pellet for 2 (frozen at -20); for the other 2, we completed the following protocol:
1) Washed with 50 mL water
2) Resuspend in 100 mL ice-cold water
3) Divided 1ml aliquots into 8 eppendorf tubes (half induced half uninduced)
4) Added 100 and 10ul to IPTG and non IPTG plates (4 total) and grew at room temperature. Froze eppendorft tubes. Tomorrow we will unfreeze them for 6 hours on ice, then 1 hour at 4C, then replate to compare colony growth - essentially a survival assay after frozen : before frozen number of viable cells (this is obviously just a test assay)
- Plated 6 samples of the remaining 2 100 mL solutions, using 10 and 100 microliters on IPTG, LB, Amp plates and non-IPTG, LB, Amp plates and stored 1 of each concentration in 0 degrees, 4 degrees, and room temperature
Tuesday - Finished and presented project defense presentation to Sackler Institute and our advisers. - Obtained TCA protein precipitation protocol.
Wednesday
Plan:
Accomplished:
Thursday
Plan:
Accomplished:
Friday
Plan:
Accomplished:
