Difference between revisions of "Team:Virginia/Experiments"

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<h2>Paracoccus denitrificans protocols</h2>
 
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<li>1. Grow Paracoccus cultures in liquid media
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<li>2. Dilute 500mL of Paracoccus denitrificans culture in 500mL SGM17 medium
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<br><li>***COLD ROOM FROM HERE***<br>
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<li>3.Harvest by centrifuge 250mL samples each in 2 four centrifuge bottles at 5000xg at 4 C. Carefully dump out supernatant.
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<li>4.Completely resuspend (shake vigorously without up & down motion to avoid liquid loss) the pellets from the previous step in all centrifuge bottles each with 10050mL of 0.5 M sucrose solution with 10% glycerol.
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<li>5. Combine the liquid(containing resuspended pellets) into one centrifuge bottle, add 200mL water to the other empty centrifuge bottle for counterbalance.
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<li>6. Centrifuge the two bottles at 5000xg at 4 C.
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<li>7. Dump out supernatant and resuspend the pellets with 100mL of 0.5 M sucrose solution with 10% glycerol. Counter-balance bottle should have 100mL of water.
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<li>8. Centrifuge the two bottles at 5000xg at 4 C.
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<li>9. Dump out supernatant and resuspend the resulting pellets in 10mL of 0.5 M sucrose solution with 10% glycerol.
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<li>10. Aliquot the final 10mL of liquid into 0.25mL(250uL) with 40 microcentrifuge tubes.
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<li>11. Flash freeze with liquid nitrogen and store at -80 C.
  
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Revision as of 20:03, 31 October 2017


Paracoccus denitrificans protocols

  • 1. Grow Paracoccus cultures in liquid media
  • 2. Dilute 500mL of Paracoccus denitrificans culture in 500mL SGM17 medium
  • ***COLD ROOM FROM HERE***
  • 3.Harvest by centrifuge 250mL samples each in 2 four centrifuge bottles at 5000xg at 4 C. Carefully dump out supernatant.
  • 4.Completely resuspend (shake vigorously without up & down motion to avoid liquid loss) the pellets from the previous step in all centrifuge bottles each with 10050mL of 0.5 M sucrose solution with 10% glycerol.
  • 5. Combine the liquid(containing resuspended pellets) into one centrifuge bottle, add 200mL water to the other empty centrifuge bottle for counterbalance.
  • 6. Centrifuge the two bottles at 5000xg at 4 C.
  • 7. Dump out supernatant and resuspend the pellets with 100mL of 0.5 M sucrose solution with 10% glycerol. Counter-balance bottle should have 100mL of water.
  • 8. Centrifuge the two bottles at 5000xg at 4 C.
  • 9. Dump out supernatant and resuspend the resulting pellets in 10mL of 0.5 M sucrose solution with 10% glycerol.
  • 10. Aliquot the final 10mL of liquid into 0.25mL(250uL) with 40 microcentrifuge tubes.
  • 11. Flash freeze with liquid nitrogen and store at -80 C.

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    Measurement protocols

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