Difference between revisions of "User:S Sibert/Notebook/SBB13Ntbk-Stephanie J Sibert"

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==4/4/13==
 
Ligations
 
6.5uL ddH2O
 
1uL Ligation buffer
 
1uL vector
 
1uL insert
 
0.5uL enzyme (last)
 
 
Comp Cells
 
1 tube cells
 
30uL KCM
 
(no ddH2O)
 
 
==4/9/13==
 
no colonies in new plate, 3 colonies in old plate will be used to proceed.
 
 
picked colonies. by ejecting pipet tip into tube.
 
 
==4/11/13==
 
1)miniprep culture<br>
 
2) restriction map EcoRI/XhoI<br>
 
6ul H2O<br>
 
1ul buffer<br>
 
2ul DNA<br>
 
0.5ul XhoI<br>
 
0.5ul EcoRI<br>
 
put in thermo cycler 11:23<br>
 
3) run a gel<br>
 
Lane 7-Sample A<br>
 
Lane 8-Sample B<br>
 
Lane 9-Sample C<br>
 
 
4) No results.<br>
 
 
==4/16/13==
 
Starting with <br>
 
50ul rxn<br>
 
31.5 ul H20<br>
 
5ul dNTP<br>
 
10ul Phusion buffer<br>
 
1ul template (sample from 3/12/13)<br>
 
1ul 10mm oligo 1 (CCOMT2-3 diluted)<br>
 
1ul 10mm oligo 2 (CCOMT2-4 diluted)<br>
 
0.5ul Phusion<br>
 
 
 
Tube placed in thermocycler labeled ccomt2 4/16/13
 
 
 
 
Golden Gate Set up:<br>
 
(n)ul ddH2O to 10ul<br>
 
1ul Ligase Buffer<br>
 
0.5ul Ligase<br>
 
0.5ul BsmB1<br>
 
1ul PER CONSTRUCT<br>
 
0.5ul vertor<br>
 
 
==4/18/2013==
 
sample processed by jc anderson to miniprep step <br>
 
miniprep<br>
 
 
==4/19/2012==
 
sequence confirmed<br>
 
golden gate<br>
 
Protocol:<br>
 
6.5ul ddH2o<br>
 
1ul ligase buffer<br>
 
0.5ul ligase<br>
 
0.5ul BsMb1<br>
 
1ul of synthon mic<br>
 
o.5ul vector<br>
 
 
==4/23/13==
 
 
Transform<br>
 
1 tube<br>
 
20ul KCM<br>
 
no h20<br>
 
Rescue <br>
 
plate<br>
 
 
==4/25/13==
 
~miniprep~<br>
 
6.5ul H20<br>
 
2ul Plasmid<br>
 
1ul NEB2<br>
 
0.5ul Bsa1<br>
 
<Br>
 
~map~<br>
 
lanes 7,8,9 <br>
 
A,B,D respectivly<br>
 
 
 
 
 
~seq~<br>
 
 
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Revision as of 14:20, 1 December 2013

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