User:S Sibert/Notebook/SBB13Ntbk-Stephanie J Sibert

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(4/2/13)
Current revision (16:20, 1 December 2013) (view source)
 
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==4/4/13==
 
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Ligations
 
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6.5uL ddH2O
 
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1uL Ligation buffer
 
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1uL vector
 
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1uL insert
 
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0.5uL enzyme (last)
 
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Comp Cells
 
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1 tube cells
 
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30uL KCM
 
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(no ddH2O)
 
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==4/9/13==
 
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no colonies in new plate, 3 colonies in old plate will be used to proceed.
 
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picked colonies. by ejecting pipet tip into tube.
 
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==4/11/13==
 
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1)miniprep culture<br>
 
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2) restriction map EcoRI/XhoI<br>
 
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6ul H2O<br>
 
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1ul buffer<br>
 
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2ul DNA<br>
 
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0.5ul XhoI<br>
 
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0.5ul EcoRI<br>
 
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put in thermo cycler 11:23<br>
 
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3) run a gel<br>
 
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Lane 7-Sample A<br>
 
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Lane 8-Sample B<br>
 
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Lane 9-Sample C<br>
 
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4) No results.<br>
 
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==4/16/13==
 
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Starting with <br>
 
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50ul rxn<br>
 
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31.5 ul H20<br>
 
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5ul dNTP<br>
 
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10ul Phusion buffer<br>
 
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1ul template (sample from 3/12/13)<br>
 
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1ul 10mm oligo 1 (CCOMT2-3 diluted)<br>
 
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1ul 10mm oligo 2 (CCOMT2-4 diluted)<br>
 
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0.5ul Phusion<br>
 
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Tube placed in thermocycler labeled ccomt2 4/16/13
 
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Golden Gate Set up:<br>
 
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(n)ul ddH2O to 10ul<br>
 
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1ul Ligase Buffer<br>
 
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0.5ul Ligase<br>
 
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0.5ul BsmB1<br>
 
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1ul PER CONSTRUCT<br>
 
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0.5ul vertor<br>
 
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==4/18/2013==
 
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sample processed by jc anderson to miniprep step <br>
 
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miniprep<br>
 
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==4/19/2012==
 
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sequence confirmed<br>
 
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golden gate<br>
 
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Protocol:<br>
 
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6.5ul ddH2o<br>
 
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1ul ligase buffer<br>
 
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0.5ul ligase<br>
 
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0.5ul BsMb1<br>
 
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1ul of synthon mic<br>
 
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o.5ul vector<br>
 
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==4/23/13==
 
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Transform<br>
 
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1 tube<br>
 
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20ul KCM<br>
 
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no h20<br>
 
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Rescue <br>
 
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plate<br>
 
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==4/25/13==
 
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~miniprep~<br>
 
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6.5ul H20<br>
 
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2ul Plasmid<br>
 
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1ul NEB2<br>
 
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0.5ul Bsa1<br>
 
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<Br>
 
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~map~<br>
 
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lanes 7,8,9 <br>
 
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A,B,D respectivly<br>
 
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~seq~<br>
 
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