GMU talk:Synthetic Biology/Bacteria Game/iGEM Wiki: Difference between revisions
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** incubation of swarm plates at room temperature for 20 h | ** incubation of swarm plates at room temperature for 20 h | ||
*** sufficient humidity was ensured by petri dishes filled with water | *** sufficient humidity was ensured by petri dishes filled with water | ||
** photo was taken (Canon EOS 5D Mark II) every 30 sec. for 6 h | ** a photo was taken (by Canon EOS 5D Mark II) every 30 sec. for 6 h | ||
* inoculation of over-night cultures from the day cultures | * inoculation of over-night cultures from the day cultures | ||
Revision as of 16:45, 4 October 2010
iGem Wiki preparation page
Introduction
Small introductory text.
Description
A in depth description of the non-technical concept.
Technical Description
The concept of the game is based on the ability of some harmless wildtype bacteria to swim in soft media. Swimming enables the microbes to consume further nutrients if those in their vicinity are already consumed. All bacteria try to get away from the starting point as fast as possible to access fresh media. This mechanism can be employed for a game setup. Selection and culturing of best swimmers leads to propagation of ideal swimming characteristics. That is, why training may help gain a competitive edge. Those bacteria can easily be stored in the fridge with the supplied materials without any risk.
The showdown competition is run by synthetic bacteria. Predators and prey communicate and regulate each other's density. Via molecular signals, the predator cells kill the prey while living prey rescues predators. The diverse and colorful crowd surrounding the spectacle was genetically engineered to carry different pigments which has been appreciated at the iGEM competition 2009.
Wetlab Notebook
- usage of E. chromi principle (pigmented E. coli cells)
- as a colorful crowd
- plasmids available in iGEM Spring 2010 DNA Distribution (see table 1)
- as a colorful crowd
- testing of a synthetic E.coli predator-prey system (Song et al., 2009), (Ballagaddé et al., 2008)
- for final assault on the swarming plate and under the microscope
- kindly provided by R. Smith (see table 2)
- for final assault on the swarming plate and under the microscope
- wildtype E. coli
- for the game-kit
- offered by A. Kern
- for the game-kit
30/09/2010
- transformation of TOP10 cells with plasmids (see table 1) out of the registry following standard recommendations
- over-night growth on selective LB agar plates (10 g Tryptone, 5 g Yeast extract, 10 g NaCl and 15 g Agar ad 1 L ddH2O + required antibiotics)
plasmid | Part | pigment color | backbone | registry location |
---|---|---|---|---|
pLA01 | Template:Part | red | Template:Part | 2010 Kit Plate 3, 6J |
pLA02 | Template:Part | orange | Template:Part | 2010 Kit Plate 3, 6N |
pLA03 | Template:Part | purple | Template:Part | 2010 Kit Plate 3, 12B |
pLA04 | Template:Part | dark green | Template:Part | 2010 Kit Plate 3, 20H |
pLA05 | Template:Part | light green | Template:Part | 2010 Kit Plate 3, 20J |
- glycerol stocks (prepared directly after arrived) from predator and prey cells (see table 2) were plated on selective LB agar plates (s. a.)
sample | name/function | cell strain | plasmid | marker |
---|---|---|---|---|
1a | predator | MG1655 | ptetLuxRLasI-luxCcdA(SC101), placCcdBs-tetGFPuv(LVA) | Cm, Kan |
3a | prey | MG1655 | pLasRLuxI-luxCcdBs, ptet-mCherry(ColE1) | Cm, Kan |
31/09/2010
- inoculation of day cultures with colonies from the previous day
- in 5 mL LB + required antibiotic
- preparation of TB soft agar plates (10 g Tryptone, 5 g NaCl and 3 g Agar ad 1 L ddH2O + required antibiotics swarm plates) for swarming
- inoculation by carefully pipetting 3 μL of liquid culture into the solidified agar
- incubation of swarm plates at room temperature for 20 h
- sufficient humidity was ensured by petri dishes filled with water
- a photo was taken (by Canon EOS 5D Mark II) every 30 sec. for 6 h
- inoculation of over-night cultures from the day cultures
01/10/2010
- preparation and conduction of microscopy experiments with predator-prey system following liquid-phase protocols
- snapshots were taken every second for one minute at beginning and at the very end of the experiments
- time-lapse took 3 h with a picture each 20 seconds using given filters for GFPuv and mCherry