From 2008.igem.org
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| + | <p>Pawel: pET15b vector transformed into TOP10 strain for plasmid purification </p> |
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Revision as of 17:47, 1 October 2008
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Cloning of protein A DNA to OmpA constructs
1. Digestion of pKS-A plasmid, pACYC177+OmpA_omega and pACYC177+OmpA_alpha with NotI and SacI (pACYC177 were also dephosporylated with CIAP)
2. Gel electrophoresis and gel-out of proper bands (420 bp - pKS-A lane, 4050 bp - pACYC177+OmpA_omega lane and 4300 bp pACYC177+OmpA_alpha lane).
3. Ligation of A DNA fragment with both pACYC177 vectors.
4. Transformation of E. coli TOP10 strain with ligations.
5. Transformants plating on LB + kanamycin.
Pawel: pET15b vector transformed into TOP10 strain for plasmid purification
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