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Team:UNICAMP-Brazil/Notebooks/October 13
From 2009.igem.org
(New page: {{:Team:UNICAMP-Brazil/inc_topo}} {{:Team:UNICAMP-Brazil/inc calendar}} __NOTOC__ ==''' YeastGuard '''== ====New strategy: pGEM==== *<p style=”text-align:justify;”>The final confi...) |
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*<p style=”text-align:justify;”>The final confirmation of the lysozyme biobrick was done by digestion of the three plasmids chosen yesterday with ''EcoR''I and ''Pst''I. The digestion confirmed two lysozyme biobricks (~600bp of the part and ~3000bp of the vector).</p> | *<p style=”text-align:justify;”>The final confirmation of the lysozyme biobrick was done by digestion of the three plasmids chosen yesterday with ''EcoR''I and ''Pst''I. The digestion confirmed two lysozyme biobricks (~600bp of the part and ~3000bp of the vector).</p> | ||
| - | + | [[Image:digestãolisozima.jpg|300px|center]] | |
*<p style=”text-align:justify;”>We did miniprep of the promoters’ biobricks and digested the plasmids with ''EcoR''I and ''Pst''I to confirm the correct insertion. The digestion of pJEN1 confirmed 1 positive colony (~1000bp) and the pDLD digestion confirmed 2 colonies (~500bp).</p> | *<p style=”text-align:justify;”>We did miniprep of the promoters’ biobricks and digested the plasmids with ''EcoR''I and ''Pst''I to confirm the correct insertion. The digestion of pJEN1 confirmed 1 positive colony (~1000bp) and the pDLD digestion confirmed 2 colonies (~500bp).</p> | ||
| - | + | [[Image:235pDLD.jpg|350px|center]] | |
*<p style=”text-align:justify;”>We digested the correct biobricks with ''Spe''I and ''Pst''I in order to open the vectors and connect the parts. The digestion showed expected fragments (pDLD: 3729bp and pJEN1: 4264bp).</p> | *<p style=”text-align:justify;”>We digested the correct biobricks with ''Spe''I and ''Pst''I in order to open the vectors and connect the parts. The digestion showed expected fragments (pDLD: 3729bp and pJEN1: 4264bp).</p> | ||
| - | + | [[Image:prepDLD.jpg|300px|center]] | |
*<p style=”text-align:justify;”>We purified the fragments from the agarose gel and connected them to lysozyme and to the reporter gene YFP, both digested with ''Xba''I and ''Pst''I. Then we transformed these ligations in competent ''E. coli'' and plated in LB+Amp media.</p> | *<p style=”text-align:justify;”>We purified the fragments from the agarose gel and connected them to lysozyme and to the reporter gene YFP, both digested with ''Xba''I and ''Pst''I. Then we transformed these ligations in competent ''E. coli'' and plated in LB+Amp media.</p> | ||
Revision as of 04:12, 21 October 2009
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