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Team:University of Lethbridge/Notebook/Project1September
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September 10, 2008
Selina, Christa, Munima
Created 10 mL sterile 100 mM theophylline solution. Aliquotted into microfuge tubes and placed in cabinet in teaching lab.
Note: higher solubility in EtOH (see Merck index) but will only reach 100 mM in a super-saturated H20 solution (heat up to 50C directly before use).
September 11, 2008
Christa, Selina, Munima
Made motility media tubes (~20 blank, ~12 with 1 mM theophylline, ~10 with 0.25 mM theophylline, 1 with 50 mM theophylline). Attempts to make 'layered' motility media stab tubes was a complete disaster.
Labeled tubes and placed in teaching lab beside LB culture tubes - DO NOT MIX THESE UP!!!!
September 12, 2008
Selina
Counted colonies from previous sucessful competency/transformation attempt (Aug. 25/08).
- CaCl2 treated DH5a + pUC19 = ~3 colonies/uL (~300 colonies in 100 uL) - RP1616 + pUC19 = ~2 colonies/uL (~ 200 colonies in 100 uL)
- CaCl2 treated DH5a + pTopp = ~1.8 colonies/uL (~450 colonies in 250 uL) - RP1616 + pTopp = ~0.2 colonies/uL (~ 20 colonies in 100 uL)
September 22, 2008
Christa, Munima, John
Objective: Assess motility with our newly made motility media (Sept. 11/08).
Stabbed tubes of 0 mM, 0.25 mM, 1 with 0.5 mM and 1 mM [theophylline] with RP1616 (from -80C glycerol stock) or DH5alpha + pTopp from Aug. 25/08 plate for negative and positive control, respectively.
Incubated at @ 37 C overnight.
September 24, 2008
Christa
Objective: Glycerol stock cells plated on August 25 & 26, 2008
Subcultured colony from RP 1616 CaCl2 + pTopp from August 26, 2008 plate (single colony) into 1- 5mL culture tube containing 100ug/mL amp. Subcultured colonies from DH5a + pTopp from August 25, 2008 plate into 2- 5mL culture tubes containg 100ug/mL amp. Left in shaker incubator at 37 C overnight.
September 25, 2008
Christa
Objective: Glycerol stock cells plated on August 25 & 26, 2008
No growth observed from the colony subcultured from RP1616 CaCl2 + pTopp. Left in shaker incubator overnight at 37 C.
Growth observed in culture tubes contain colonies from DH5a + pTopp. Made 4 glycerol stocks of these tubes, stored in the Wieden - 80 C labelled iGEM DH5a + pTopp September 25/08.
