IGEM:UW/2008/Notebook/iGEM UW Team/2008/06/03: Difference between revisions
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|style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> University of Warsaw iGEM Team 2008</span> | |||
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | |||
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1. Transformants E.coli TOP10 with pZC320 (liquid LB) | |||
2. Gel electrophoresis (PCR products) and isolation from proper bands | |||
3. Digestion with HindIII and SalI | |||
4. Digestion of pMPM-T5 + AID with HindIII and SalI | |||
5. Clean-up products of p.3 and p.4 reactions | |||
6. Ligation of p.5 products | |||
7. Transformation of E.coli TOP10 with ligation product | |||
8. Transformants onto LB+tetracycline | |||
9. Digestion of pMPM-T5 + transcription fusion with EcoRHI and HindIII | |||
10. Use of T4 polimerase for blunting | |||
11. Gel electrophoresis | |||
12. Isolation from proper band | |||
13. Ligation of product p.12 | |||
14. Transformation of E.coli TOP10 with ligation product | |||
15. Transformants onto LB + tetracycline | |||
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Revision as of 02:38, 11 June 2008
University of Warsaw iGEM Team 2008 | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
1. Transformants E.coli TOP10 with pZC320 (liquid LB) 2. Gel electrophoresis (PCR products) and isolation from proper bands 3. Digestion with HindIII and SalI 4. Digestion of pMPM-T5 + AID with HindIII and SalI 5. Clean-up products of p.3 and p.4 reactions 6. Ligation of p.5 products 7. Transformation of E.coli TOP10 with ligation product 8. Transformants onto LB+tetracycline 9. Digestion of pMPM-T5 + transcription fusion with EcoRHI and HindIII 10. Use of T4 polimerase for blunting 11. Gel electrophoresis 12. Isolation from proper band 13. Ligation of product p.12 14. Transformation of E.coli TOP10 with ligation product 15. Transformants onto LB + tetracycline
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