Team:Exeter/lab book/3gip/wk2

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         <a href="https://2012.igem.org/Team:Exeter/lab_book/3gip/wk11"; style="color:#1d1d1b">17th - 21st September</a>
         <a href="https://2012.igem.org/Team:Exeter/lab_book/3gip/wk11"; style="color:#1d1d1b">17th - 21st September</a>
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        <a href="https://2012.igem.org/Team:Exeter/Results/inducible"; style="color:#e30614" target="_blank"><font size="3"><b>Results</b></font></a>
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</font>
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     <!--End Project Division Week Hyperlinks-->
     <!--End Project Division Week Hyperlinks-->
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   <!------------INSERT WEEKLY IMAGE HERE------------>
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<p><i><b>Afternoon</b></i></p>
<p><i><b>Afternoon</b></i></p>
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<p>•<u> Transformation:</u></p>  
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<p>•<a href="https://2012.igem.org/Team:Exeter/lab_book/proto/1" style="color:#57b947"><u>Transformation</u></a></p>  
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RBS Biobrick BBa_B0034, 2012 Distribution Kit Plate 1 2M
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<p>RBS Biobrick BBa_B0034, 2012 Distribution Kit Plate 1 2M</p>
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Tetr_rbs Biobrick BBa_J13002, 2012 Distribution Kit Plate 1 13B
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<p>Tetr_rbs Biobrick BBa_J13002, 2012 Distribution Kit Plate 1 13B</p>
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Using Invitrogen TOP10 competent cells split into 2 eppendorfs containing 25μl each.
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<p>Using Invitrogen TOP10 competent cells split into 2 eppendorfs containing 25μl each. </p>
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Spread on two Ampicillin plates for each transformation using 20μl and 100μls respectively.  
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<p>Spread on two ampicillin plates for each transformation using 20μl and 100μls respectively. </p><br>
 +
 
<p><b><u>Tuesday 17th July</u></b></p>
<p><b><u>Tuesday 17th July</u></b></p>
<p><i><b>Afternoon</b></i></p>
<p><i><b>Afternoon</b></i></p>
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<p>•<u>Adding cultures</u></p>   
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<p>•<a href="https://2012.igem.org/Team:Exeter/lab_book/proto/2" style="color:#57b947"><u>Transferring colonies to liquid medium</u></a></p>   
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Cells containing BBa_B0034 and BBa_J13002 were added into liquid medium and incubated overnight
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<p>Cells containing BBa_B0034 and BBa_J13002 were added into liquid medium and incubated overnight</p>
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Protocol was followed using ampicillin for all BioBrick parts as the selection antibiotic.
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<p>Protocol was followed using ampicillin for all BioBrick parts as the selection antibiotic. </p><br>
 +
 
<p><b><u>Wednesday 18th July</u></b></p>
<p><b><u>Wednesday 18th July</u></b></p>
<p><i><b>Morning</b></i></p>
<p><i><b>Morning</b></i></p>
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The incubator had stopped over night and so growth overnight was not optimised. During the miniprep this morning, the cultures were centrifuged with pipette tips in the tubes. The pellets were therefore very small and so the process was repeated overnight using fresh cultures.
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<p>The incubator stopped unexpectedly during the night, this meant growth was not optimised. During the miniprep this morning, the cultures were centrifuged with pipette tips in the tubes. The pellets were very small and therefore the process was repeated overnight using fresh cultures. </p><br>
 +
 
<p><i><b>Afternoon</b></i></p>
<p><i><b>Afternoon</b></i></p>
-
<p>•<u> Adding cultures</u></p>   
+
<p>•<a href="https://2012.igem.org/Team:Exeter/lab_book/proto/2" style="color:#57b947"><u>Transferring colonies to liquid medium</u></a></p>   
-
Cells containing BBa_B0034 and BBa_J13002 were added into liquid medium and incubated overnight
+
<p>Cells containing BBa_B0034 and BBa_J13002 were added into liquid medium and incubated overnight. </p>
-
Protocol was followed using ampicillin for all BioBrick parts as the selection antibiotic.
+
<p>Protocol was followed using ampicillin for all BioBrick parts as the selection antibiotic. </p><br>
-
<p><b><u>Thursday 19th July</u></b></p>
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 +
<b><u>Thursday 19th July</u></b></p>
<p><i><b>Morning</b></i></p>
<p><i><b>Morning</b></i></p>
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<p>•<u> Mini-Prepping</u> of BBa_B0034 and BBa_J13002</p>
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<p>•<a href="http://www.fermentas.com/templates/files/tiny_mce/coa_pdf/coa_k0502.pdf" style="color:#57B947" target="_blank"><u>Miniprepping</u></a> of BBa_B0034 and BBa_J13002</p>
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Both BBa_B0034 and BBa_J13002 were nanodropped and recorded low concentrations. This was put down to inexperience with minipreps.
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Both BBa_B0034 and BBa_J13002 were nanodropped and recorded low concentrations. This was put down to inexperience with mini-preps. </p><br>
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<p>•<u> Gel Electrophoresis</u> on a 2% gel was run to check fragment sizes of BBa_B0034 and BBa_J13002 using the EcoR1 and Pst1 enzymes
+
<p>•<u> Gel Electrophoresis</u> on a 2% gel was run to check fragment sizes of BBa_B0034 and BBa_J13002 using the EcoR1 and Pst1 enzymes. </p>
-
BBa_B0034 showed no band but is only 13bp large and BBa_J13002 showed a very faint band.  
+
<p>BBa_B0034 showed no band but is only 13bp large and BBa_J13002 showed a very faint band. </p>
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    <p><u>Website Designed and Built by: Ryan Edginton, James Lynch & Alex Clowsley</u> &nbsp;&nbsp;|&nbsp;&nbsp;
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    <a href="https://igem.org/Team.cgi?id=764" style="color:#57B947" target="_blank"><u>Contact Us</u></a>  &nbsp;&nbsp;|&nbsp;&nbsp;
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Latest revision as of 00:04, 27 September 2012

ExiGEM2012 Lab Book 3GP wk2

The 3-Gene Inducible Plasmid: 16th - 20th July 2012

 Monday 16th July

Afternoon

•Transformation

RBS Biobrick BBa_B0034, 2012 Distribution Kit Plate 1 2M

Tetr_rbs Biobrick BBa_J13002, 2012 Distribution Kit Plate 1 13B

Using Invitrogen TOP10 competent cells split into 2 eppendorfs containing 25μl each.

Spread on two ampicillin plates for each transformation using 20μl and 100μls respectively.


Tuesday 17th July

Afternoon

•Transferring colonies to liquid medium

Cells containing BBa_B0034 and BBa_J13002 were added into liquid medium and incubated overnight

Protocol was followed using ampicillin for all BioBrick parts as the selection antibiotic.


Wednesday 18th July

Morning

The incubator stopped unexpectedly during the night, this meant growth was not optimised. During the miniprep this morning, the cultures were centrifuged with pipette tips in the tubes. The pellets were very small and therefore the process was repeated overnight using fresh cultures.


Afternoon

•Transferring colonies to liquid medium

Cells containing BBa_B0034 and BBa_J13002 were added into liquid medium and incubated overnight.

Protocol was followed using ampicillin for all BioBrick parts as the selection antibiotic.


Thursday 19th July

Morning

•Miniprepping of BBa_B0034 and BBa_J13002

Both BBa_B0034 and BBa_J13002 were nanodropped and recorded low concentrations. This was put down to inexperience with mini-preps.


• Gel Electrophoresis on a 2% gel was run to check fragment sizes of BBa_B0034 and BBa_J13002 using the EcoR1 and Pst1 enzymes.

BBa_B0034 showed no band but is only 13bp large and BBa_J13002 showed a very faint band.

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