Team:Hawaii/Notebook/2008-06-13

From 2008.igem.org

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(→Quote of the Day)
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===Prepped Cells for Glycerol Stock/Seed Stock===
===Prepped Cells for Glycerol Stock/Seed Stock===
:<strong> Krystle</strong>
:<strong> Krystle</strong>
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Inoculated a single colony in each 10 mL LB + antibiotic<br>
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:* Inoculated a single colony in each 10 mL LB + antibiotic<br>
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<br>
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{| border="1"
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<table style="text-align: left; width: 478px; height: 205px;"
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|+ '''Title'''
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border="1" cellpadding="2" cellspacing="2">
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!Antibiotic
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  <tbody>
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!Cell type
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    <tr>
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!Plasmid
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      <td style="font-weight: bold; text-align: center;">Antibiotic</td>
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|-
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      <td style="font-weight: bold; text-align: center;">Cell
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| rowspan="4" align="center"|Amp<sub>100</sub>||align="center"| StrataClone supercompetent cells || align="center"|pUC18  
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Type</td>
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|-
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      <td style="font-weight: bold; text-align: center;">Plasmid</td>
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|  align="center"|DH5&alpha;|| align="center"|GFP  
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    </tr>
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|-
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    <tr>
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|  align="center"|DH5&alpha;|| align="center"|tet+GFP
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      <td style="text-align: center;" colspan="1"
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|-
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rowspan="4">Amp<small><small>100</small></small></td>
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|  align="center"|DH5&alpha;|| align="center"|RBS (BBa_B0034)
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      <td style="text-align: center;">supercompetent cells</td>
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|-
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      <td style="text-align: center;">pUC18</td>
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| align="center"|Kan<sub>50</sub>||align="center"|HB101|| align="center"|RK1
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    </tr>
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|-
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    <tr>
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| align="center"|Sm<sub>2.5</sub> + Sp <sub>2.5</sub>|| align="center"|DH5&alpha;|| align="center"|pRL1383a
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      <td style="text-align: center;">DH5alpha</td>
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|}
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      <td style="text-align: center;">GFP</td>
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    </tr>
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    <tr>
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      <td style="text-align: center;">DH5alpha</td>
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      <td style="text-align: center;">tet+GFP</td>
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    </tr>
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    <tr>
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      <td style="text-align: center;">DH5alpha</td>
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      <td style="text-align: center;">RBS</td>
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    </tr>
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    <tr>
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      <td style="text-align: center;">Kan<small><small>50</small></small></td>
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      <td style="text-align: center;">HB101</td>
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      <td style="text-align: center;">RK1</td>
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    </tr>
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    <tr>
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      <td style="text-align: center;">Sm<small><small>2.5</small></small>
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+ Sp <small><small>2.5</small></small></td>
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      <td style="text-align: center;">DH5alpha</td>
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      <td style="text-align: center;">pRL1383a</td>
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    </tr>
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  </tbody>
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===Prepped Cells for Competency===
===Prepped Cells for Competency===
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:* (2) 300 mL batches of LB + 1.5% Agar. Stored in 4C.
:* (2) 300 mL batches of LB + 1.5% Agar. Stored in 4C.
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== Drylab Work ==
 
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= Discussion =
= Discussion =
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= Quote of the Day =
= Quote of the Day =
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[[Image:DSC00746.JPG | ''E. coli'' homes. Enough said.]]
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[[Image:DSC00746.JPG |thumb|left|225px| ''E. coli'' homes. Enough said.]]
{{Team:Hawaii/Footer}}
{{Team:Hawaii/Footer}}
__NOTOC__
__NOTOC__

Revision as of 18:15, 14 June 2008

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Things we did today

Wetlab work

Prepped Cells for Glycerol Stock/Seed Stock

Krystle
  • Inoculated a single colony in each 10 mL LB + antibiotic
Title
Antibiotic Cell type Plasmid
Amp100 StrataClone supercompetent cells pUC18
DH5αGFP
DH5αtet+GFP
DH5αRBS (BBa_B0034)
Kan50HB101RK1
Sm2.5 + Sp 2.5DH5αpRL1383a


Prepped Cells for Competency

Grace
  • Checked yesterday's inoculation. OD600=1.2 (too high). Need to redo.
  • Made SOB for new inoculation.
  • Inoculated 250 μl SOB with 100 μl DH5α. Inoculated 250 μl SOB with 10 μl DH5α. Incubated overnight at 27C with shaking (150 rpm).

Cell density test

Grace
  • Retrieved plates from SC incubator. All plates had too many colonies (lawn). Need to redo.
  • Made BG-11 plates for retest next week.

Made LB agar stock

Grace
  • (2) 300 mL batches of LB + 1.5% Agar. Stored in 4C.

Discussion

  • We need to make more BG-ll 50x stock solution and TES 50x stock solution
  • Kelly from SC lab will be in 12pm-3pm. We should retrieve our triparental conjugation plates then.
  • Grace, Krystle, and Norman will come in at 9am tomorrow to plate triparental conjugates, make competent cells, and glycerol stock purified Biobrick colonies.

Quote of the Day

E. coli homes. Enough said.


[http://manoa.hawaii.edu/ Sponsor_UHM.gif][http://manoa.hawaii.edu/ovcrge/ Sponsor_OVCRGE.gif][http://www.ctahr.hawaii.edu Sponsor_CTAHR.gif]


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