Team:Hawaii/Notebook/2008-06-14
From 2008.igem.org
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Things we did today
Wetlab Work
Cell density test for PCC6803
- Krystle
- Plated 10-5, 10-7, 10-8 dilutions of PCC6803 (OD750 = .071 and 1.15) on BG-11
- Plates in SC incubator
Glycerol stocked Biobrick colonies
- Krystle
- Combined 0.4ml 80% glycerol w/ 0.8ml of transformed E. coli grown overnight in LB + antibiotic
- Cryostock could not be made for pUC18 because of contamination in overnight culture
Made competent cells
- Grace
- Same protocol as 6/2 competent cells
- OD600 = 0.4567 (overnight culture)
- OD600 = 1.1-1.2 (after CCMB80 added)
Triparental conjugation
- Grace
- Streaked BG-11+sp+sm plates with conjugates
- Incubated in light & CO2 @ 30C in SC incubator
Discussion
For competent cells, should cool eppendorf tray at 4C. After aliquot, transfer tubes to -80C EtOH tray and freeze at -80C. GP suggests we aliquot with tubes in dry ice.
Was Callahan lab hood working?
Quote of the Day
- "Yeah, I'm trying to stave off the real world by staying in school a little longer." - KS
- "Ooooooooooooooooooooooooh" -KS, "YEAH Krystle, it's shiny." -GK
[http://manoa.hawaii.edu/ ][http://manoa.hawaii.edu/ovcrge/ ][http://www.ctahr.hawaii.edu ]