From 2008.igem.org
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| - | <h3>Michał</h3>
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| - | <ol><li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation of plasmids</a> from cultures inocluated on <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/4_August_2008">previous day</a>. </li>
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| - | <li> Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of isolated plasmids with BamHI and SacI. </li></ol>
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Revision as of 22:13, 11 October 2008
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- Digest of pACYC177+OmpA_omega_deltaA_alpha (from 25 July) with BamHI and NotI. DNA ends blunting with Klenow fragment (3 hr).
- Gel elctrophoresis and gel-out of proper band - 4300 bp.
- Ligation of isolated DNA fragment (1 hr).
- Transformationof E. coli TOP10 strain with ligation.
- Transformants plating on LB + kanamycin.
Checking for presence of A on the cell membrane
Piotr
- Pouring bacteria by drops onto nitrocellulose
- Drying
- Blocking
- Anti-A antibody binding
- Washing
- Anti-rabbit antibody binding
- Developing with BCIP and NBT
[photo of the gel is to be placed here]
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