Revathireddy (Talk | contribs) |
Revathireddy (Talk | contribs) |
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<h2>Week 11: Sequencing results arrive</h2> | <h2>Week 11: Sequencing results arrive</h2> | ||
− | <p style="font-family: 'Lato'; font-size:15px"> Sequencing results were positive only for <i>glnA</i>. We anyway decided to get biobricked <i>glnA</i> synthesized from IDT. Since the PR in our kitplate is without a rbs, we decided to get that too synthesized from IDT. We sent out the sequences for the same. We decided to have <i>glnA</i>, <i>gltB</i>, <i>gltD</i> along with PR in one plasmid and BBa_K1604022 with BBa_I74211 in another plasmid containing a different ori in the same cell. We planned to carry out a 3A assembly using BBa_1604022, BBa_I74211 and BBa_JO4450. | + | <p style="font-family: 'Lato'; font-size:15px"> Sequencing results were positive only for <i>glnA</i>. We anyway decided to get biobricked <i>glnA</i> and <i>gltD</i> synthesized from IDT. Since the PR in our kitplate is without a rbs, we decided to get that too synthesized from IDT. We sent out the sequences for the same. We decided to have <i>glnA</i>, <i>gltB</i>, <i>gltD</i> along with PR in one plasmid and BBa_K1604022 with BBa_I74211 in another plasmid containing a different ori in the same cell. We planned to carry out a 3A assembly using BBa_1604022, BBa_I74211 and BBa_JO4450. |
<p></p> | <p></p> | ||
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<h2>Week 12: 3A assembly!</h2> | <h2>Week 12: 3A assembly!</h2> | ||
− | <p style="font-family: 'Lato'; font-size:15px"> Sequencing results were positive only for <i>glnA</i> We carried out a variant of 3A assembly using BBa_1604022 (cut with EcoRI and SpeI), BBa_I74211(cut with XbaI and PstI) and BBa_JO4450 (cut with EcoRI and PstI). Unfortunately, it did not work. | + | <p style="font-family: 'Lato'; font-size:15px"> Sequencing results were positive only for <i>glnA</i> We carried out a variant of 3A assembly using BBa_1604022 (cut with EcoRI and SpeI), BBa_I74211(cut with XbaI and PstI) and BBa_JO4450 (cut with EcoRI and PstI). Unfortunately, it did not work.<br> |
− | + | Meanwhile, we received our gblocks for <i>glnA</i>, <i>gltD</i>and rbs+PR from IDT. The concentrations of each of these were found to be extremely low that we reordered all the theree gblocks again. <br> | |
+ | <img src= "https://static.igem.org/mediawiki/parts/3/3a/ICT-Mumbai_notebook_gelimage3.png"> <br> | ||
+ | From left to right: <i>glnA</i> fragment 1 (G1_F1), <i>glnA</i> fragment 2 (G2_F2), <i>gltD</i> fragment 1 (G3_F1), <i>gltD</i> fragment 2 (G3_F2), rbs+PR follwed by 1kb DNA ladder. </p> | ||
+ | <p></p> | ||
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Revision as of 02:20, 1 November 2017