Team:Yale/Our Project/Notebook/Week 4

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Each reaction was let to run 20 minutes at room temperature before being transformed into Xl1-Blue cells according to the standard <a href="http://2010.igem.org/Team:Yale/Our_Project/Protocols/transformation"> transformation protocol </a> and then plated on ampicillin LB plates and let to grow overnight.<br/>
Each reaction was let to run 20 minutes at room temperature before being transformed into Xl1-Blue cells according to the standard <a href="http://2010.igem.org/Team:Yale/Our_Project/Protocols/transformation"> transformation protocol </a> and then plated on ampicillin LB plates and let to grow overnight.<br/>
<b> Cultures for miniprep & glycerol stock redo </b> <br/>
<b> Cultures for miniprep & glycerol stock redo </b> <br/>
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In preparation for potential ligation failure, once again inoculated liquid cultures of Amp LB  for miniprep with pSB74,P0312,R0011, B0015, and J23114 transformants, making two 5 mL cultures for each plasmid.  When OD reached 0.6, once again made glycerol stocks from 0.5 mL of each solution and 0.5 mL of filter-sterilized 50% glycerol.  This time flash-froze the stocks immediately, rather than waiting overnight as.  <br/>
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In preparation for potential ligation failure, once again inoculated liquid cultures of Amp LB  for miniprep with pSB74,P0312,R0011, B0015, and J23114 transformants, making two 5 mL cultures for each plasmid.  When OD reached 0.6, once again made glycerol stocks from 0.5 mL of each solution and 0.5 mL of filter-sterilized 50% glycerol.  This time flash-froze the stocks immediately, rather than waiting overnight as on <a href="http://2010.igem.org/Team:Yale/Our_Project/Notebook/Week_3" 6/23.</a> <br/>

Revision as of 11:09, 27 October 2010

iGEM Yale

lab notebook: week 4 (6/28 -7/4)

  • Monday 6/28--PCR amplification of thiosulfate reductase operon phsABC
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  • Tuesday 6/29--Gel extraction of PCR amplified phs, TSI agar slant assay for hydrogen sulfide production, & pre-ligation double digestion
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  • Wednesday 6/30--Troubleshooting PCR amplification of thiosulfate reductase gene & continued work toward ligation
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  • Thursday short content
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  • Fridays short content
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  • Saturday short content
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  • Sunday short content
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