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Team:Warsaw/Calendar-Main/3 June 2008
From 2008.igem.org
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| - | <p>1. | + | <p>1. DNA (PCR products) gel electrophoresis and isolation of DNA from proper bands<br> |
| - | 2. | + | 2. Digestion of DNA with HindIII and SalI<br> |
| - | 3. | + | 3. Digestion of pMPM-T5 + AID with HindIII and SalI<br> |
| - | 4. Clean-up products of p.2 and p.3 reactions<br> | + | 4. Clean-up of products of p.2 and p.3 reactions<br> |
5. Ligation of p.4 products (pMPM-T5 + AID transcription fusion with AID-T7 RNA-polymerase translation fusion)<br> | 5. Ligation of p.4 products (pMPM-T5 + AID transcription fusion with AID-T7 RNA-polymerase translation fusion)<br> | ||
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6. Transformation of E.coli TOP10 with ligation product<br> | 6. Transformation of E.coli TOP10 with ligation product<br> | ||
| - | 7. | + | 7. Plating transformants on LB+tetracycline<br> |
| - | 8. | + | 8. Digestion of pMPM-T5 + transcription fusion with EcoRHI and HindIII<br> |
9. Use of T4 polymerase for blunting<br> | 9. Use of T4 polymerase for blunting<br> | ||
| - | 10. | + | 10. DNA gel electrophoresis<br> |
| - | 11. Isolation from proper band<br> | + | 11. Isolation of DNA from proper band<br> |
12. Ligation of product p.11 (pMPM-T5 + T7 RNA-polymerase)<br> | 12. Ligation of product p.11 (pMPM-T5 + T7 RNA-polymerase)<br> | ||
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13. Transformation of E.coli TOP10 with ligation product<br> | 13. Transformation of E.coli TOP10 with ligation product<br> | ||
| - | 14. | + | 14. Plating transformants on LB + tetracycline</p> |
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<img src="https://static.igem.org/mediawiki/2008/a/a4/UW_AAT.jpg" width="500"/> | <img src="https://static.igem.org/mediawiki/2008/a/a4/UW_AAT.jpg" width="500"/> | ||
Revision as of 13:28, 26 September 2008
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1. DNA (PCR products) gel electrophoresis and isolation of DNA from proper bands
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