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Team:Grinnell/Notebook/Gels/DspB
From 2011.igem.org
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<a name='20110718_BBa_OR_PrsaA%2BoptdspB%2BrsaA_pMR10' href='https://2011.igem.org/File:20110718_BBa_OR_PrsaA%2BoptdspB%2BrsaA_pMR10.jpg'> | <a name='20110718_BBa_OR_PrsaA%2BoptdspB%2BrsaA_pMR10' href='https://2011.igem.org/File:20110718_BBa_OR_PrsaA%2BoptdspB%2BrsaA_pMR10.jpg'> | ||
<img alt ='20110718_BBa_OR_PrsaA%2BoptdspB%2BrsaA_pMR10' src='https://static.igem.org/mediawiki/2011/5/5a/20110718_BBa_OR_PrsaA%2BoptdspB%2BrsaA_pMR10.jpg' /> | <img alt ='20110718_BBa_OR_PrsaA%2BoptdspB%2BrsaA_pMR10' src='https://static.igem.org/mediawiki/2011/5/5a/20110718_BBa_OR_PrsaA%2BoptdspB%2BrsaA_pMR10.jpg' /> | ||
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| + | </td> | ||
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| + | <a name='20110719_Pxyl%2BodspB%2BrsaA_re-do_c1,3,4,5,6_std' href='https://2011.igem.org/File:20110719_Pxyl%2BodspB%2BrsaA_re-do_c1,3,4,5,6_std.jpg'> | ||
| + | <img alt ='20110719_Pxyl%2BodspB%2BrsaA_re-do_c1,3,4,5,6_std' src='https://static.igem.org/mediawiki/2011/4/46/20110719_Pxyl%2BodspB%2BrsaA_re-do_c1%2C3%2C4%2C5%2C6_std.jpg' /> | ||
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| + | <a name='20110720_Pxyl%2BodspB%2BrsaA_re-do_c7-14_WTdspB_c5' href='https://2011.igem.org/File:20110720_Pxyl%2BodspB%2BrsaA_re-do_c7-14_WTdspB_c5.jpg'> | ||
| + | <img alt ='20110720_Pxyl%2BodspB%2BrsaA_re-do_c7-14_WTdspB_c5' src='https://static.igem.org/mediawiki/2011/3/30/20110720_Pxyl%2BodspB%2BrsaA_re-do_c7-14_WTdspB_c5.jpg' /> | ||
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| + | <a name='20110722_BBa%2BodspB%2BrsaA%2BpMR10_c1,5-9' href='https://2011.igem.org/File:20110722_BBa%2BodspB%2BrsaA%2BpMR10_c1,5-9.jpg'> | ||
| + | <img alt ='20110722_BBa%2BodspB%2BrsaA%2BpMR10_c1,5-9' src='https://static.igem.org/mediawiki/2011/a/aa/20110722_BBa%2BodspB%2BrsaA%2BpMR10_c1%2C5-9.jpg' /> | ||
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Lane 1: ladder; lanes 2-5: transformants that should carry BBa_K081005, optimized <I>dspB</i>, and <i>rsaA</i> in pMR10; lanes 6-9: transformants that should carry P<sub>rsaA</sub>, optimized <I>dspB</i>, and <i>rsaA</i> in pMR10; lane 10: standard DNA fragment of optimized <i>dspB</i> with <i>rsaA</i>. Lane 7 carries the desired gene sequence. | Lane 1: ladder; lanes 2-5: transformants that should carry BBa_K081005, optimized <I>dspB</i>, and <i>rsaA</i> in pMR10; lanes 6-9: transformants that should carry P<sub>rsaA</sub>, optimized <I>dspB</i>, and <i>rsaA</i> in pMR10; lane 10: standard DNA fragment of optimized <i>dspB</i> with <i>rsaA</i>. Lane 7 carries the desired gene sequence. | ||
| + | </td> | ||
| + | <td> | ||
| + | Gel of PCR of transformants that should carry P<sub>xyl</sub>, optimized <i>dspB</i>, and <i>rsaA</i>. Lane 1: ladder; lanes 2-6: PCR products; lane 7: standard fragment of <i>dspB</i> with <i>rsaA</i>. | ||
| + | </td> | ||
| + | <td> | ||
| + | Gel of PCR of transformants that should carry P<sub>xyl</sub>, optimized <i>dspB</i>, and <i>rsaA</i>. Lane 1: ladder; lanes 2-9: transformant PCR products; lane 10: digested miniprep of what should have been WT <i>dspB</i>, but showed up as the wrong fragment size. | ||
| + | </td> | ||
| + | <td> | ||
| + | PCR of transformants that should carry BBa_K081005, optimized <i>dspB</i>, and <i>rsaA</i> in pMR10. Lane 1: ladder; lanes 2-7: PCR products. | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td> | ||
| + | <a name='20110722_Pxyl%2BodspB%2BrsaA%2BpMR10' href='https://2011.igem.org/File:20110722_Pxyl%2BodspB%2BrsaA%2BpMR10.jpg'> | ||
| + | <img alt ='20110722_Pxyl%2BodspB%2BrsaA%2BpMR10' src='https://static.igem.org/mediawiki/2011/f/f1/20110722_Pxyl%2BodspB%2BrsaA%2BpMR10.jpg' /> | ||
| + | </a> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td> | ||
| + | PCR of transformants that should carry P<sub>xyl</sub>, optimized <i>dspB</i>, and <i>rsaA</i> in pMR10. Lane 1: ladder; lanes 2-8: PCR products. | ||
</td> | </td> | ||
</table> | </table> | ||
</html> | </html> | ||
Revision as of 16:36, 25 July 2011
Gels for DspB
2011.07.03-2011.07.09
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| Gel proving the successful transformation of dspB containing plasmid into E. coli Top10. Lane 1: ladder; lanes 2-4: digests (EcoRI and PstI) of miniprep DNA from overnights of transformants. There is a clear band at the appropriate size for dspB in all of the experimental lanes. | Gel showing dspB with various promoter inserts as contrasted to a standard dspB with no promoter. Lane 1: dspB (WT) + PrsaA; lane 2: with Pxyl; lanes 3,4: with BBa_K081005; lane 5: standard dspB; lane 6: ladder. Not all transformants show successful insertion of the desired promoter, but it is clear which were successful when compared to the standard dspB. | Gel of transformants that should contain both dspB and a promoter insert. Lane 1: ladder; lanes 2-4: dspB with PrsaA; lanes 5-7: dspB with Pxyl; lane 8: standard dspB with no promoter. | Gel of more transformants of dspB with Pxyl. Lane 1: ladder; lanes 2-5: transformants; lane 6: standard dspB with no promoter. |
2011.07.10-2011.07.16
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| Gel of transformants with synthesized dspB and esp with codons optimized for expression in Caulobacter after ligation into pSB1C3. Lane 1: ladder; lanes 2-5: optimized dspB; lanes 6-9: optimized esp. Lane 4 shows successful insertion of dspB into pSB1C3, but none of the optimized esp transformants show the desired insert. | Gel of colony PCR of transformants that should carry WT dspB behind the Pxyl promoter. Lanes 1-4: colonies 9, 10, 12, and 15 from transformation plate; lane 5: dspB WT standard from PCR. | Gel for extraction of WT dspB behind Pxyl. Lane 1: ladder; lanes 2,3: miniprep digest. | Gel of colony PCR of transformants carrying optimized dspB and rsaA C-terminal. Lane 1: ladder; lanes 2-5: transformation colonies 1-4; lane 6: dspB standard from PCR of WT dspB. |
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| Lane 1: ladder; lanes 2-5: transformants that should carry PrsaA, optimized dspB, and rsaA; lanes 6-9: transformants that should carry Pxyl, optimized dspB, and rsaA; lane 10: standard optimized dspB from PCR. None of these transformants showed success. | Lane 1: ladder; lanes 2-5: transformants that should carry Pxyl, optimized dspB, and rsaA; lane 6: standard optimized dspB with rsaA from PCR. None of these colonies were successful. | Lane 1: standard optimized dspB with rsaA from PCR; lanes 2,3: transformants that should carry WT dspB behind Pxyl; lane 4: ladder. No successful transformations. | Lane 1: ladder; lanes 2-5: transformants that should carry BBa_K081005, optimized dspB, and rsaA; lane 6: standard optimized dspB with rsaA fragment. Lanes 3 and 5 carry the desired combination of genes. |
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| Lane 1: ladder; lanes 2-7: transformants that should carry Pxyl, optimized dspB, and rsaA; lane 8: standard fragment of optimized dspB with rsaA. None of the transformants carried the desired gene sequence. |
2011.07.17-2011.07.23
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| Lane 1: ladder; lanes 2-5: transformants that should carry BBa_K081005, optimized dspB, and rsaA in pMR10; lanes 6-9: transformants that should carry PrsaA, optimized dspB, and rsaA in pMR10; lane 10: standard DNA fragment of optimized dspB with rsaA. Lane 7 carries the desired gene sequence. | Gel of PCR of transformants that should carry Pxyl, optimized dspB, and rsaA. Lane 1: ladder; lanes 2-6: PCR products; lane 7: standard fragment of dspB with rsaA. | Gel of PCR of transformants that should carry Pxyl, optimized dspB, and rsaA. Lane 1: ladder; lanes 2-9: transformant PCR products; lane 10: digested miniprep of what should have been WT dspB, but showed up as the wrong fragment size. | PCR of transformants that should carry BBa_K081005, optimized dspB, and rsaA in pMR10. Lane 1: ladder; lanes 2-7: PCR products. |
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| PCR of transformants that should carry Pxyl, optimized dspB, and rsaA in pMR10. Lane 1: ladder; lanes 2-8: PCR products. |
