Team:ETH Zurich/lab/bead

From 2014.igem.org

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(Production)
(Production)
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===Production===
===Production===
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Bacteria are resuspended and diluted in NaCl-solution (0.9 % in H<sub>2</sub>O) so as to achieve the desired cell density. Here, we aimed at a comparably high concentration of 10<sup>7</sup> bacteria per bead. The resuspension was added to alginate (2.5%) to reach a alginate concentration of 2%. The viscous solution is filled into a syringe containing an appropriate needle device and extruded at a constant, slow velocity. Ideally, the droplets should fall form a height of approximately 30 cm into a 100 mM CaCl<sub>2</sub> solution. In the CaCl<sub>2</sub> solution gelling of the alginate droplets will occur instantaneously. To avoid excessive salt stress for the bacteria the beads should be transferred to a 10 mM CaCl<sub>2</sub> solution.
 
[[File:ETH2014_BeadsProduction.jpg|left|300px]]
[[File:ETH2014_BeadsProduction.jpg|left|300px]]
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Bacteria are resuspended and diluted in NaCl-solution (0.9 % in H<sub>2</sub>O) so as to achieve the desired cell density. Here, we aimed at a comparably high concentration of 10<sup>7</sup> bacteria per bead. The resuspension was added to alginate (2.5%) to reach a alginate concentration of 2%. The viscous solution is filled into a syringe containing an appropriate needle device and extruded at a constant, slow velocity. Ideally, the droplets should fall form a height of approximately 30 cm into a 100 mM CaCl<sub>2</sub> solution. In the CaCl<sub>2</sub> solution gelling of the alginate droplets will occur instantaneously. To avoid excessive salt stress for the bacteria the beads should be transferred to a 10 mM CaCl<sub>2</sub> solution.
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A more detailed protocol can be found [https://2014.igem.org/Team:ETH_Zurich/lab/protocols here.]
A more detailed protocol can be found [https://2014.igem.org/Team:ETH_Zurich/lab/protocols here.]
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Revision as of 16:29, 15 October 2014

iGEM ETH Zurich 2014