Team:Yale/Our Project/Notebook/Week 4

From 2010.igem.org

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Sunday 7/4--Ligation attempt #2 transformants
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<h4> Transformed Colonies from Ligation Attempt #2</h4>
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<li>Checked on plated transformants and observed the following: </li>
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<b>phsABC/B0015 system</b><br/>
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Reaction #1 (control) 2 colonies<br/>
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Reaction #2 (2:1 insert:vector) 4 colonies, numbered 1B-4B<br/>
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Reaction #3 (2:1 insert:vector) 5 colonies, numbered 5B-9B<br/>
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Reaction #4 (large control) 0 colonies<br/>
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<b>J23114/P0312 system </b> <br/>
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Reaction #5 (control) numerous* colonies <br/>
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Reaction #6 (2:1 insert:vector) numerous colonies, somewhat more than #5<br/>
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Reaction #7 (3:1 insert:vector) numerous colonies,about double #5<br/>
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Reaction #8 (5:1 insert:vector) numerous colonies, about double #5<br/>
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numerous=more than readily countable<br/>
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<li>Inoculated 5 mL liquid Amp/LB cultures with colonies 1B-9B and let grow overnight at 37 on shaker </li>
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<li> Set aside plates 6-8 in the fridge to work with later. </li>
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<i>This day's activities are also recorded on page 46 in the hard copy of the lab notebook. </i>
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Revision as of 11:55, 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 7/1--Smelly bacteria (yay!) & ongoing ligation efforts
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  • Fridays 7/2--Confirmation of hydrogen sulfide production plus more ligation work
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  • Saturday 7/3--More attempts to put a terminator on thiosulfate reductase and a promoter on lacI
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  • Sunday 7/4--Ligation attempt #2 transformants
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