User:Daniel Ramirez/Notebook/UNAM Genomics Mexico 2011/2011/05/24
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χρόνος πέρασμα May 24th 2011
a) The digested plasmid, blunted and with the PCR amplification product, all this treated with ligase. Expected outcome: Red colonies; this means the E.coli that did grew contain a plasmid that confers gentamicin resistance (pBBRMCS5 backbone) and the amplification product that has the RFP gene.
b) The digested plasmid, blunted and dephosphorilated, all this treated with ligase. Expected outcome: No colonies; even when it was treated with ligase, there shouldn't be any circularized plasmid as two DNA tips can't be joined if they are dephosphorilated. So no gentamicin resistance gene is available for the bacteria to survive.
c) The digested plasmid and blunted, all this treated with ligase. Expected outcome: Grey normal colonies, the pBBRMCS5 plasmid, without the RFP gene, should be recircularized, each bacteria containing this plasmid should survive.
d) An empty petri dish. Expected outcome: No colonies; as the solid medium was added with gentamicin, no bacteria should be able to survive, unless they possess some sort of resistance to the antibiotic.