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Team:Weimar-Heidelberg Arts/Project/Bacteria Game/Wetlab
From 2010.igem.org
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| - | + | <p><a href="https://2010.igem.org/Team:Weimar-Heidelberg_Arts/Project/Bacteria_Game">Return to the main Bacteria Game page</a> | |
| - | + | <h2> <span class="mw-headline" id="Wetlab_Notebook"> Wetlab Notebook </span></h2> | |
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<ul><li> usage of <a href="https://2009.igem.org/Team:Cambridge" class="external text" rel="nofollow">E. chromi principle</a> (pigmented <i>E. coli</i> cells) | <ul><li> usage of <a href="https://2009.igem.org/Team:Cambridge" class="external text" rel="nofollow">E. chromi principle</a> (pigmented <i>E. coli</i> cells) | ||
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</li></ul> | </li></ul> | ||
| - | <h3 | + | <h3><span class="mw-headline" id="30.2F09.2F2010"> 30/09/2010 </span></h3> |
<ul><li> <a href="https://2010.igem.org/Transformation" class="external text" rel="nofollow">transformation</a> of TOP10 cells with plasmids (see table 1) out of the registry following <a href="http://partsregistry.org/Help:Spring_2010_DNA_distribution#DNA_Kit_Plate_Instructions" class="external text" rel="nofollow">standard recommendations</a> | <ul><li> <a href="https://2010.igem.org/Transformation" class="external text" rel="nofollow">transformation</a> of TOP10 cells with plasmids (see table 1) out of the registry following <a href="http://partsregistry.org/Help:Spring_2010_DNA_distribution#DNA_Kit_Plate_Instructions" class="external text" rel="nofollow">standard recommendations</a> | ||
</li><li> 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 ddH<sub>2</sub>O + required antibiotics) | </li><li> 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 ddH<sub>2</sub>O + required antibiotics) | ||
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| - | <h3 | + | <h3> <span class="mw-headline" id="01.2F10.2F2010"> 01/10/2010 </span></h3> |
<ul><li> inoculation of day cultures with colonies from the previous day | <ul><li> inoculation of day cultures with colonies from the previous day | ||
<ul><li> in 5 mL LB + required antibiotic | <ul><li> in 5 mL LB + required antibiotic | ||
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</li><li> inoculation of over-night cultures from the day cultures | </li><li> inoculation of over-night cultures from the day cultures | ||
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<ul><li> preparation and conduction of microscopy experiments with predator-prey system following <a href="http://www.nature.com/nchembio/journal/v5/n12/extref/nchembio.244-S1.pdf" class="external text" rel="nofollow">liquid-phase protocols</a> | <ul><li> preparation and conduction of microscopy experiments with predator-prey system following <a href="http://www.nature.com/nchembio/journal/v5/n12/extref/nchembio.244-S1.pdf" class="external text" rel="nofollow">liquid-phase protocols</a> | ||
Latest revision as of 17:31, 27 October 2010
Return to the main Bacteria Game page
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 | BBa_K274110 | red | pSB1A2 | 2010 Kit Plate 3, 6J |
| pLA02 | BBa_K274210 | orange | pSB1A2 | 2010 Kit Plate 3, 6N |
| pLA03 | BBa_K274002 | purple | pSB1T3 | 2010 Kit Plate 3, 12B |
| pLA04 | BBa_K274003 | dark green | pSB1K3 | 2010 Kit Plate 3, 20H |
| pLA05 | BBa_K274004 | light green | pSB1K3 | 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 |
01/10/2010
- 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
02/10/2010
- 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
