Hello,
Can anyone help me understand what my results are showing. My aim was to express the SucC gene in pET15B from pET3a-SucC. I performed a colony PCR with 4 ligation tubes and 1 control and ran the cells through a agarose gel. I loaded 10μl of DNA ladder and then loaded the cells in the gel and this is my result:
Your question really needs more information. What is in each well? What size bands are you expecting and observing?
Did you do a transformation and get cells after your ligations then run PCR on the colonies or did you literally run PCR on your ligations?
Ben, it is still not clear what you did. You said you did a transformation and then ran a PCR. Do you mean that you picked one colony from each plate, did a miniprep on it, then did a PCR on the miniprep (or a colony PCR on a single colony). For example your lanes 1 and 5 look like mini prepped plasmid DNA but 2-4 look like PCR product. Are the primers such that you would expect to see a 5700 bp product from the wrong clone and the 1500 from the right clone?
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