"
Team:Warsaw/Calendar-Main/23 May 2008
From 2008.igem.org
MKrzyszton (Talk | contribs) |
MKrzyszton (Talk | contribs) |
||
| Line 6: | Line 6: | ||
<p>Michał K.:</p> | <p>Michał K.:</p> | ||
| - | <p>1. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of PCR product for transcription fusion (T7 RNA-polimerase) with HindIII and SalI</p> | + | <p>1. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of PCR product for transcription fusion (T7 RNA-polimerase) with HindIII and SalI (2x Tango buffer).</p> |
<p>2. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of PCR product for translation fusion (AID+T7 RNA-polimerase) with NcoI and XbaI</p> | <p>2. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of PCR product for translation fusion (AID+T7 RNA-polimerase) with NcoI and XbaI</p> | ||
| Line 12: | Line 12: | ||
<p>3. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">Clean-up</a> of digested products from p.1 and p.2</p> | <p>3. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">Clean-up</a> of digested products from p.1 and p.2</p> | ||
| - | <p>4. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of pMPM-T5+AID with HindIII and SalI</p> | + | <p>4. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of pMPM-T5+AID with HindIII and SalI (2x Tango buffer).</p> |
<p>5. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> with NcoI and XbaI</p> | <p>5. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> with NcoI and XbaI</p> | ||
Revision as of 22:43, 30 September 2008
 |
|
||||||||
 |
|
||||||||
Michał K.: 1. Digestation of PCR product for transcription fusion (T7 RNA-polimerase) with HindIII and SalI (2x Tango buffer). 2. Digestation of PCR product for translation fusion (AID+T7 RNA-polimerase) with NcoI and XbaI 3. Clean-up of digested products from p.1 and p.2 4. Digestation of pMPM-T5+AID with HindIII and SalI (2x Tango buffer). 5. Digestation of pMPM-T5 with NcoI and XbaI 6. Gel-electrophresis and gel-out of proper bands. 7. Ligation of purified products from p.1 and p.4 8. Electroporation of E.coli TOP10 with ligation product 9. Ligation of digested products from p.4 and p.5 10. Electroporation of E.coli TOP10 with ligation product 11. Transformants plating on LB + tetracycline http://openwetware.org/images/thumb/6/61/UW_A%2BT.jpg/350px-UW_A%2BT.jpg http://openwetware.org/images/thumb/c/c7/UW_AT.jpg/350px-UW_AT.jpg
|
|
