Agarose gel of EcoRI-digested putative recombinant plasmid from the cloning of the PCR fragments:
Miniprep DNAs from the white colonies were digested with EcoRI, and the enzyme digestion was then run on an 0.8% agarose gel in TBE buffer. Lane 1, 1 kb ladder; lanes 2 and 3, miniprep DNAs from two white colonies from the Top A PCR fragment ligations; lanes 4 and 5, miniprep DNAs from two white colonies from the cp PCR fragment ligations; lane 6, 1 kb ladder. The very top band (slowest moving) for lanes 2, 3, 4 and 5 is non-digested recombinant plasmid. The recombinant plasmid in lane 3 was named pElios1 (Top A insert) and will be sequenced.
Miniprep DNAs were prepared from four colonies from the transformation plates for Top A inserts and cp inserts. All four colonies for the Top A PCR fragment had inserts of the correct size and three of the four colonies for the cp PCR fragment had inserts of the correct size.
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