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Team:Warsaw/Calendar-Main/13 May 2008
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<li>Setup of separate cultures from colonies of transformants pMPMT5+AID (liquid LB+tetracycline).</li> | <li>Setup of separate cultures from colonies of transformants pMPMT5+AID (liquid LB+tetracycline).</li> | ||
| - | <li> Optimization of conditions for <html><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> to obtain AID DNA fragment - annealing temperature gradient (from 62°C to 82°C). Fig. 1.<br> | + | <li> Optimization of conditions for <html><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> to obtain AID DNA fragment - annealing temperature gradient (from 62°C to 82°C). <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/13_May_2008#fig1">Fig. 1</a>.<br> |
Primers: | Primers: | ||
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| - | <li> Optimization of conditions for <html><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - T7 RNA polymerase for translational fusion; annealing temperature gradient (from 62°C to 82°C). Fig. 2.<br> | + | <li> Optimization of conditions for <html><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - T7 RNA polymerase for translational fusion; annealing temperature gradient (from 62°C to 82°C). <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/13_May_2008#fig1">Fig. 2</a>.<br> |
Primers: | Primers: | ||
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</li> | </li> | ||
| - | <li> Optimization of conditions for <html><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - T7 RNA polymerase for transcriptional fusion; annealing temperature gradient (from 62°C to 82°C). Fig. 2. <br> | + | <li> Optimization of conditions for <html><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - T7 RNA polymerase for transcriptional fusion; annealing temperature gradient (from 62°C to 82°C). <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/13_May_2008#fig1">Fig. 2</a>. <br> |
Primers: | Primers: | ||
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| - | Template DNA: <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> (negative control - Fig. 3.) genomic DNA<br> | + | Template DNA: <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> (negative control - <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/13_May_2008#fig1">Fig. 3</a>.) genomic DNA<br> |
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</p> | </p> | ||
| - | <img src="https://static.igem.org/mediawiki/2008/7/71/Aid_pcr_grad_WAW.jpg" width=350/><var><b>Fig. 1.</b> Gradient PCR products for AID: 1-DNA ladder; 2 to 13-PCR products-border annealing temperatures of gradient: 62°C (lane 2) to 82°C (lane 13)</var> | + | <a name="fig1"><img src="https://static.igem.org/mediawiki/2008/7/71/Aid_pcr_grad_WAW.jpg" width=350/></a><var><b>Fig. 1.</b> Gradient PCR products for AID: 1-DNA ladder; 2 to 13-PCR products-border annealing temperatures of gradient: 62°C (lane 2) to 82°C (lane 13)</var> |
| - | <img src="https://static.igem.org/mediawiki/2008/9/97/T7grad_WAW1.jpg" width=350/> | + | <a name="fig2"><img src="https://static.igem.org/mediawiki/2008/9/97/T7grad_WAW1.jpg" width=350/></a> |
<var><b>Fig. 2.</b> Gradient PCR products: upper - for transcriptional fusion, lower - for translation fusion; 1-DNA ladder; 2 to 12-PCR products-border annealing temperatures of gradient: 62°C (lane 2) to 82°C (lane 12)</var> | <var><b>Fig. 2.</b> Gradient PCR products: upper - for transcriptional fusion, lower - for translation fusion; 1-DNA ladder; 2 to 12-PCR products-border annealing temperatures of gradient: 62°C (lane 2) to 82°C (lane 12)</var> | ||
| - | <img src="https://static.igem.org/mediawiki/2008/6/6c/T7grad_WAW2.jpg" width=350/> | + | <a name="fig3"><img src="https://static.igem.org/mediawiki/2008/6/6c/T7grad_WAW2.jpg" width=350/></a> |
<var><b>Fig. 3.</b> Gradient PCR products for negative control (Top10 genomic DNA): 1-DNA ladder; 2 to 6-PCR products-border annealing temperatures of gradient: 62°C (lane 2) to 82°C (lane 6).</var> | <var><b>Fig. 3.</b> Gradient PCR products for negative control (Top10 genomic DNA): 1-DNA ladder; 2 to 6-PCR products-border annealing temperatures of gradient: 62°C (lane 2) to 82°C (lane 6).</var> | ||
Revision as of 23:05, 11 October 2008
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Preparation of pMPMT5+AID construct and PCRs for fusionsMichał K.
 Fig. 1. Gradient PCR products for AID: 1-DNA ladder; 2 to 13-PCR products-border annealing temperatures of gradient: 62°C (lane 2) to 82°C (lane 13)
Fig. 2. Gradient PCR products: upper - for transcriptional fusion, lower - for translation fusion; 1-DNA ladder; 2 to 12-PCR products-border annealing temperatures of gradient: 62°C (lane 2) to 82°C (lane 12)
Fig. 3. Gradient PCR products for negative control (Top10 genomic DNA): 1-DNA ladder; 2 to 6-PCR products-border annealing temperatures of gradient: 62°C (lane 2) to 82°C (lane 6).
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