Team:Grinnell/Notebook/Gels/Esp

From 2011.igem.org

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Gel of colony PCR of transformants that should carry BBa_K081005, WT <i>esp</i>, and <i>rsaA</i> in pMR10, P<sub>xyl</sub>, WT <i>esp</i>, and <i>rsaA</i> in pSB1C3, or P<sub>rsaA</sub>, WT <i>esp</i>, and <i>rsaA</i> in pSB1C3. Lane 1: ladder; lanes 2-5: BBa_K081005, WT <i>esp</i>, and <i>rsaA</i> in pMR10; lanes 6,7: P<sub>xyl</sub>, WT <i>esp</i>, and <i>rsaA</i> in pSB1C3; lanes 8,9: P<sub>rsaA</sub>, WT <i>esp</i>, and <i>rsaA</i> in pSB1C3.  Transformant in lane 4 was successful.
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Gel of colony PCR of transformants that should carry BBa_K081005, WT <i>esp</i>, and <i>rsaA</i> in pMR10, P<sub>xyl</sub>, WT <i>esp</i>, and <i>rsaA</i> in pSB1C3, or P<sub>rsaA</sub>, WT <i>esp</i>, and <i>rsaA</i> in pSB1C3. Lane 1: ladder; lanes 2-5: BBa_K081005, WT <i>esp</i>, and <i>rsaA</i> in pMR10; lanes 6,7: P<sub>xyl</sub>, WT <i>esp</i>, and <i>rsaA</i> in pSB1C3; lanes 8,9: P<sub>rsaA</sub>, WT <i>esp</i>, and <i>rsaA</i> in pSB1C3.  
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<a name='20110718_optesprsaAGXIDTSMART_optesprsaAC3' href='https://2011.igem.org/File:20110718_optesprsaAGXIDTSMART_optesprsaAC3.jpg'>
<a name='20110718_optesprsaAGXIDTSMART_optesprsaAC3' href='https://2011.igem.org/File:20110718_optesprsaAGXIDTSMART_optesprsaAC3.jpg'>
<img alt ='20110718_optesprsaAGXIDTSMART_optesprsaAC3' src='https://static.igem.org/mediawiki/2011/3/34/20110718_optesprsaAGXIDTSMART_optesprsaAC3.jpg' />
<img alt ='20110718_optesprsaAGXIDTSMART_optesprsaAC3' src='https://static.igem.org/mediawiki/2011/3/34/20110718_optesprsaAGXIDTSMART_optesprsaAC3.jpg' />
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<a name='20110720_PrsaA%2Boesp%2BrsaA_oesp%2BrsaAC3' href='https://2011.igem.org/File:20110720_PrsaA%2Boesp%2BrsaA_oesp%2BrsaAC3.jpg'>
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<img alt='20110720_PrsaA%2Boesp%2BrsaA_oesp%2BrsaAC3' src='https://static.igem.org/mediawiki/2011/5/5f/20110720_PrsaA%2Boesp%2BrsaA_oesp%2BrsaAC3.jpg' />
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Lane 1: ladder; lanes 2-7: transformants that should carry optimized <i>esp</i> with <i>rsaA</i> from gel extraction in IDTSMART-AMP. Lanes 3, 4, and 6 have the desired gene sequence.
Lane 1: ladder; lanes 2-7: transformants that should carry optimized <i>esp</i> with <i>rsaA</i> from gel extraction in IDTSMART-AMP. Lanes 3, 4, and 6 have the desired gene sequence.
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PCR of transformants that should carry P<sub>rsaA</sub>, optimized <i>esp</i>, and <i>rsaA</i> or optimized <i>esp</i> with <i>rsaA</i> in pSB1C3. Lane 1: ladder; lanes 2-7:combo with P<sub>rsaA</sub>; lanes 8-10: combo without promoter. Lanes 3, 8, and 10 all have the desired inserts.
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<a name='20110720_Pxyl%2Boesp%2BrsaA_oesp%2BrsaAC3' href='https://2011.igem.org/File:20110720_Pxyl%2Boesp%2BrsaA_oesp%2BrsaAC3.jpg'>
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<img alt='20110720_Pxyl%2Boesp%2BrsaA_oesp%2BrsaAC3' src='https://static.igem.org/mediawiki/2011/8/84/20110720_Pxyl%2Boesp%2BrsaA_oesp%2BrsaAC3.jpg' />
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<a name='20110722_Bba%2Boesp' href='https://2011.igem.org/File:20110722_Bba%2Boesp.jpg'>
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<img alt='20110722_Bba%2Boesp' src='https://static.igem.org/mediawiki/2011/6/6a/20110722_Bba%2Boesp.jpg' />
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<a name='20110723_BBa%2Boesp' href='https://2011.igem.org/File:20110723_BBa%2Boesp.jpg'>
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<img alt='20110723_BBa%2Boesp' src='https://static.igem.org/mediawiki/2011/7/73/20110723_BBa%2Boesp.jpg' />
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<a name='20110723_PrsaA%2Boesp%2BrsaA%2BpMR10' href='https://2011.igem.org/File:20110723_PrsaA%2Boesp%2BrsaA%2BpMR10.jpg'>
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<img alt='20110723_PrsaA%2Boesp%2BrsaA%2BpMR10' src='https://static.igem.org/mediawiki/2011/8/87/20110723_PrsaA%2Boesp%2BrsaA%2BpMR10.jpg' />
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Lane 1: ladder; lanes 2-7: PCR of transformants that should carry P<sub>xyl</sub>, optimized <i>esp</i>, and <i>rsaA</i>; lanes 8-10: transformants that should carry optimized <i>esp</i> with <i>rsaA</i> in pSB1C3.
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Lane 1: ladder; lanes 2-7: transformants that should carry BBa_K081005 and optimized <i>esp</i>.
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Gel of digests of miniprepped overnight liquid culture of cells that should carry BBa_K081005 and optimized <i>esp</i>. Lane 1: ladder; lanes 2-10: digests.
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Lane 1: ladder; lanes 2-7: PCR of transformants that should carry P<sub>rsaA</sub>, optimized <i>esp</i>, and <i>rsaA</i> in pMR10. Lane 4 was successful.
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<a name='20110723_Pxyl%2Boesp%2BrsaA%2BpMR10' href='https://2011.igem.org/File:20110723_Pxyl%2Boesp%2BrsaA%2BpMR10.jpg'>
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<img alt='20110723_Pxyl%2Boesp%2BrsaA%2BpMR10' src='https://static.igem.org/mediawiki/2011/d/d3/20110723_Pxyl%2Boesp%2BrsaA%2BpMR10.jpg' />
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Lane 1: ladder; lanes 2-7: PCR of transformants that should carry P<sub>xyl</sub>, optimized <i>esp</i>, and <i>rsaA</i> in pMR10.
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==2011.07.24-2011.07.31==
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<a name='20110725_Pxyl%2Boesp%2BrsaA%2BpMR10' href='https://2011.igem.org/File:20110725_Pxyl%2Boesp%2BrsaA%2BpMR10.jpg'>
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<img alt='20110725_Pxyl%2Boesp%2BrsaA%2BpMR10' src='https://static.igem.org/mediawiki/2011/2/23/20110725_Pxyl%2Boesp%2BrsaA%2BpMR10.jpg' />
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<a name='20110727_BBa%2Boesp%2BrsaA' href='https://2011.igem.org/File:20110727_BBa%2Boesp%2BrsaA.jpg'>
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<img alt='20110727_BBa%2Boesp%2BrsaA' src='https://static.igem.org/mediawiki/2011/3/35/20110727_BBa%2Boesp%2BrsaA.jpg' />
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<a name='20110728_BD1_Caulobacter' href='https://2011.igem.org/File:20110728_BD1_Caulobacter.jpg'>
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<a name='20110729_BBa%2Boesp%2BrsaA%2BpMR10_C3' href='https://2011.igem.org/File:20110729_BBa%2Boesp%2BrsaA%2BpMR10_C3.jpg'>
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<img alt='20110729_BBa%2Boesp%2BrsaA%2BpMR10_C3' src='https://static.igem.org/mediawiki/2011/a/ac/20110729_BBa%2Boesp%2BrsaA%2BpMR10_C3.jpg' />
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Lane 1: ladder; lanes 2-8: digests of transformants that should contain P<sub>xyl</sub>, optimized <i>esp<i>, and <i>rsaA</i>.
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Lane 1: ladder; lanes 2-8: digests of miniprep of transformants that should carry BBa_K081005, optimized <i>esp</i>, and <i>rsaA</i>. No success.
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Lane 1: ladder; lanes 2-5: PCR of <i>Caulobacter</i> colonies believed to carry BBa_K081005, WT <i>esp</i>, and <i>rsaA</i>. All passed.
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Lane 1: ladder; lanes 2,3: PCR of transformants that should carry BBa_K081005, optimized <i>esp</i>, and <i>rsaA</i> in pMR10; lanes 4-10: PCR of transformants that should carry BBa_K081005, optimized <i>esp</i>, and <i>rsaA</i> in pSB1C3. No success.
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</table></html>
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==2011.08.01-2011.08.07==
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<html><table class="gallery" frame='void' rules='none'>
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<a name='20110801_BBa%2Boesp%2BrsaA%2BpMR10' href='https://2011.igem.org/File:20110801_BBa%2Boesp%2BrsaA%2BpMR10.jpg'>
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<img alt='20110801_BBa%2Boesp%2BrsaA%2BpMR10' src='https://static.igem.org/mediawiki/2011/0/0d/20110801_BBa%2Boesp%2BrsaA%2BpMR10.jpg' />
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Lane 1: ladder; lanes 2-10: transformants that should contain BBa_K081005, optimized <i>esp</i>, and <i>rsaA</i>. Lanes 2 and 8 show success.
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</table></html>

Latest revision as of 21:32, 1 August 2011

Grinnell Menubar

Gels for Esp

2011.06.05-2011.06.11

2011.06.12-2011.06.18

2011.06.19-2011.06.25

2011.06.26-2011.07.02

2011.07.03-2011.07.09

2011.07.10-2011.07.16

2011.07.17-2011.07.23

2011.07.24-2011.07.31

2011.08.01-2011.08.07