Team:Hawaii/Notebook/2008-06-13

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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|375px| ''E. coli'' homes. Enough said.]][[Image:DSC00747.JPG|thumb|right|225px|Krystle & Grace confusing Norman, again. Two halves of the same person?]]
{{Team:Hawaii/Footer}}
{{Team:Hawaii/Footer}}
__NOTOC__
__NOTOC__

Latest revision as of 23:11, 14 June 2008

Projects Events Resources
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Notebook (t) Meetings (t)

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.
Krystle & Grace confusing Norman, again. Two halves of the same person?


[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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