Revathireddy (Talk | contribs) |
Revathireddy (Talk | contribs) |
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<h2>Week 17: indC arrives!</h2> | <h2>Week 17: indC arrives!</h2> | ||
<p style="font-family: 'Lato'; font-size:15px"> We received indC (BBa_K1152008, BBa_K1152018 and BBa_K1152019) from the iGEM registry, whereas BBa_K1152013 and pRB5 were generously sent by the Heidelberg team. BBa_K1152008 was transformed, prepped, ligated with pET 43.1a+<i>glnA</i> and transformed in <i>E. coli</i> DH5-α competent cells. <br> | <p style="font-family: 'Lato'; font-size:15px"> We received indC (BBa_K1152008, BBa_K1152018 and BBa_K1152019) from the iGEM registry, whereas BBa_K1152013 and pRB5 were generously sent by the Heidelberg team. BBa_K1152008 was transformed, prepped, ligated with pET 43.1a+<i>glnA</i> and transformed in <i>E. coli</i> DH5-α competent cells. <br> | ||
+ | Four colonies of transformants (43+glnA+indC) were screened by performing RE digestion with NdeI<br> | ||
+ | Results: Agarose gel electrophoresis revealed band of ~7000bp in all colonies tested.<br> | ||
+ | To reconfirm, colony 3 (C-3) was inoculated in LB+Amp. Plasmid prep of this colony was performed the next day and confirmed by double digestion with NotI and XhoI. Release of indC fragment (3855bp) was checked by running a 1% agarose gel. We could not see a band of the correct size.<br> | ||
+ | <img src="https://static.igem.org/mediawiki/parts/5/5e/ICT-Mumbai_notebook_gelimage2.png"> <br> | ||
+ | Lane 2 contains 1kb DNA ladder whereas lane 3 contains RE digested pET 43.1a+<i>glnA</i>+<i>indC</i>. | ||
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+ | </div> | ||
+ | </div> | ||
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+ | <div class="container right"> | ||
+ | <div class="content"> | ||
+ | <h2>Week 18: indC diaries !</h2> | ||
+ | <p style="font-family: 'Lato'; font-size:15px"> We carried out a PCR of BBa_K1152008 and ligated with pET 43.1a, BBa_K1152018 and BBa_K1152019) from the iGEM registry, whereas BBa_K1152013 and pRB5 were generously sent by the Heidelberg team. BBa_K1152008 was transformed, prepped, ligated with pET 43.1a+<i>glnA</i> and transformed in <i>E. coli</i> DH5-α competent cells. <br> | ||
Four colonies of transformants (43+glnA+indC) were screened by performing RE digestion with NdeI<br> | Four colonies of transformants (43+glnA+indC) were screened by performing RE digestion with NdeI<br> | ||
Results: Agarose gel electrophoresis revealed band of ~7000bp in all colonies tested.<br> | Results: Agarose gel electrophoresis revealed band of ~7000bp in all colonies tested.<br> |
Revision as of 20:24, 31 October 2017