Team:Lethbridge/Notebook/Lab Work/August
From 2010.igem.org
(→Aug 9/2010) |
(→Aug 9/2010) |
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<tr><td>mRBS-TetR Colony 3 | <tr><td>mRBS-TetR Colony 3 | ||
</table><br> | </table><br> | ||
- | |||
<b>Objective:</b> To determine which of the previous ligations worked.<br> | <b>Objective:</b> To determine which of the previous ligations worked.<br> | ||
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<tr><td>PstI</td><td>0.25</td></tr> | <tr><td>PstI</td><td>0.25</td></tr> | ||
</table> | </table> | ||
+ | |||
Unrestricted Control | Unrestricted Control | ||
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DNA was restricted for 1 hour at 37<sup>o</sup>C.<br> | DNA was restricted for 1 hour at 37<sup>o</sup>C.<br> | ||
- | Analyzed results on a 2% agarose gel. Load order as follows:<br> | + | Analyzed results on a 2% agarose gel with 2 (µL) of loading dye and 10 (µL) of pDNA. . Load order as follows:<br> |
+ | |||
+ | <table border ="3"> | ||
+ | <tr><td><b>Lane</b><td><b>Contents</b><td> | ||
+ | <tr><td>1<td>1kb ladder<td> | ||
+ | <tr><td>2<td>50 bp ladder<td> | ||
+ | <tr><td>3<td>dT-pTet 1 DRD<td> | ||
+ | <tr><td>4<td>dT-pTet 1 SRD<td> | ||
+ | <tr><td>5<td>dT-pTet 1 URD<td> | ||
+ | <tr><td>6<td>dT-pTet 3 DRD<td> | ||
+ | <tr><td>7<td>dT-pTet 3 SRD<td> | ||
+ | <tr><td>8<td>dT-pTet 3 URD<td> | ||
+ | <tr><td>9<td>pLacI-mRBS 1 DRD<td> | ||
+ | <tr><td>10<td>pLacI-mRBS 1 SRD<td> | ||
+ | <tr><td>11<td>pLacI-mRBS 1 URD<td> | ||
+ | <tr><td>12<td>pLacI-mRBS 2 DRD<td> | ||
+ | <tr><td>13<td>pLacI-mRBS 2 SRD<td> | ||
+ | <tr><td>14<td>pLacI-mRBS 2 URD<td> | ||
+ | <tr><td>15<td>pLacI-sRBS 1 DRD<td> | ||
+ | <tr><td>16<td>pLacI-sRBS 1 SRD<td> | ||
+ | <tr><td>17<td>pLacI-sRBS 1 URD<td> | ||
+ | <tr><td>18<td>pLacI-sRBS 3 DRD<td> | ||
+ | <tr><td>19<td>pLacI-sRBS 3 SRD<td> | ||
+ | <tr><td>20<td>pLacI-sRBS 3 URD<td> | ||
+ | </table><br> | ||
+ | |||
+ | <table border ="3"> | ||
+ | <tr><td><b>Lane</><td><b>Contents</b><td> | ||
+ | <tr><td>1<td>1 kb ladder<td> | ||
+ | <tr><td>2<td>50 bp ladder<td> | ||
+ | <tr><td>3<td>pBAD-mRBS 1 DRD<td> | ||
+ | <tr><td>4<td>pBAD-mRBS 1 SRD<td> | ||
+ | <tr><td>5<td>pBAD-mRBS 1 URD<td> | ||
+ | <tr><td>6<td>pBAD-mRBS 2 DRD<td> | ||
+ | <tr><td>7<td>pBAD-mRBS 2 SRD<td> | ||
+ | <tr><td>8<td>pBAD-mRBS 2 URD<td> | ||
+ | <tr><td>9<td>pBAD-sRBS 1 DRD<td> | ||
+ | <tr><td>10<td>pBAD-sRBS 1 SRD<td> | ||
+ | <tr><td>11<td>pBAD-sRBS 1 URD<td> | ||
+ | <tr><td>12<td>pBAD-sRBS 2DRD<td> | ||
+ | <tr><td>13<td>pBAD-sRBS 2 SRD<td> | ||
+ | <tr><td>14<td>pBAD-sRBS 2 URD<td> | ||
+ | <tr><td>15<td>mRBS-TetR 1 DRD<td> | ||
+ | <tr><td>16<td>mRBS-TetR 1 SRD<td> | ||
+ | <tr><td>17<td>mRBS-TetR 1 URD<td> | ||
+ | <tr><td>18<td>mRBS-TetR 3 DRD<td> | ||
+ | <tr><td>19<td>mRBS-TetR 3 SRD<td> | ||
+ | <tr><td>20<td>mRBS-TetR 3 URD<td> | ||
+ | </table><br> | ||
+ | |||
+ | ---- | ||
+ | <b>Objective:</b> To ligate: pBAD-sRBS/mRBS, sRBS/mRBS-TetR, TetR-dT, dT-pTet, pLacI-sRBS/mRBS, Mms6-ptet28(a), dT-PSB1C3<br> | ||
+ | |||
+ | <b>Method:</b> [[Team:Lethbridge/Notebook/Protocols|Ligation of Plasmid DNA]] | ||
+ | 15 (µL) pDNA in plasmid, and 15 (µL) of pDNA biobrick)<br> | ||
+ | |||
+ | |||
Ran gel at 100V for 40 minutes.<br> | Ran gel at 100V for 40 minutes.<br> |