From 2008.igem.org
(Difference between revisions)
|
|
| Line 17: |
Line 17: |
| | </ol></p> | | </ol></p> |
| | | | |
| - | <br>
| + | |
| - | <h3>Cloning of protein A DNA to pET15b+Omp-alfa plasmid<br>Antoni</h3>
| + | |
| - | <p><ol>
| + | |
| - | <li> Isolation of plasmids from cultures inocluated on previous day (pGeneart+A and pET15b+OmpA-alfa). </li>
| + | |
| - | <li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> pET15b+OmpA_alpha and pGeneart+A with NdeI and NotI, pET15b+OmpA_alpha<a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing"> dephosphorylation with CIAP</a> </li> </li>
| + | |
| - | <li> Gel electrophoresis of digested plasmids and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper bands </li>
| + | |
| - | <li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of pET15b+alfa and A </li>
| + | |
| - | </li>
| + | |
| - | </ol></p>
| + | |
| | | | |
| | | | |
Revision as of 23:34, 2 October 2008
 |
|
 |
|
Cloning of protein A DNA to OmpA constructs
- Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega and pACYC177+OmpA_A_alpha. Primers used:
AL+SacI and AP+NotI
- Confirmed transformant colonies (found only on pACYC177+OmpA_A_alpha plate) inoculated to liquid LB with kanamycin.
Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA
Paweł
- Ligation transformed into TOP10 strain and plated on LB+kanamycin
|
|
"
