Team:Lambert GA/labnotebook

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                 <li><a href="https://2013hs.igem.org/Team:Lambert_GA/activities" accesskey="6" title="">Activities</a></li>               
                 <li><a href="https://2013hs.igem.org/Team:Lambert_GA/activities" accesskey="6" title="">Activities</a></li>               
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<font size=4><center><b><u><h1>Lab Notebook</h1></u></b></center></font>
<font size=4><center><b><u><h1>Lab Notebook</h1></u></b></center></font>
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<th>Date</th>
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<td>June 14th</td>
<td>June 14th</td>
<td>We recorded the results of the plates. There were no colonies from the 6/12 ligations. Pictures of the 6/13 ligations have been included.
<td>We recorded the results of the plates. There were no colonies from the 6/12 ligations. Pictures of the 6/13 ligations have been included.
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<center><b>1:1 Ratio</b></center>
 
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<center><b>1:1 Ratio</b>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<b>1:2 Ratio</b>
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<br><img src="https://static.igem.org/mediawiki/2013hs/8/85/June1411.png" width="25%">
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<center><img src="https://static.igem.org/mediawiki/2013hs/c/c7/June1412.png" width="25%"></center>
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<center>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<b>1:3 Ratio</b>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<b>Antarctic Phosphatase (Just backbone)</b>
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<br><center><b>Control R0011</b></center>
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1:3 Ratio
 
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[[File:June1413.png]]
 
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Antarctic Phosphatase (Just backbone)
 
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[[File:June14Antph.png]]
 
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Control R0011
 
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[[File:June14Control.png]]
 
We did colony PCR using the HybB promers for 16 colonies, choosing the ligation mixture that was a 2:1 ratio of HybB to backbone. We set the thermocycler to 30 cycles annealing 50 degrees. Our elongation was 30 seconds. We did a diagnostic gel with 16 colonies and analyzed our results. Unfortunately, our colony PCR failed. We do not know the reason for the failure, but we suspect that
We did colony PCR using the HybB promers for 16 colonies, choosing the ligation mixture that was a 2:1 ratio of HybB to backbone. We set the thermocycler to 30 cycles annealing 50 degrees. Our elongation was 30 seconds. We did a diagnostic gel with 16 colonies and analyzed our results. Unfortunately, our colony PCR failed. We do not know the reason for the failure, but we suspect that
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Then we miniprepped the liquid cultures. We found concentrations of DNA range from 79.5ng/ul to 114.5ng/ul. Then they were digested with EcoR 37C for 15 minutes. We ran diagnostic gel on the digests.  We were expecting bands at 2070 bp for just PSB1C3 and 2463 for Colonies which contained HybB promoter sequence.  We also ran them with a control of R0011.   
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Then we miniprepped the liquid cultures. We found concentrations of DNA range from 79.5ng/ul to 114.5ng/ul. Then they were digested with EcoR 37C for 15 minutes. We ran diagnostic gel on the digests.  We were expecting bands at 2070 bp for just PSB1C3 and 2463 for Colonies which contained HybB promoter sequence.  We also ran them with a control of R0011.
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[[File:June18Gel.png]]
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Latest revision as of 03:49, 22 June 2013