Approximately 583 base pairs of sequence one was highlighted and analyzed using the linear map of the plasmid (Figure 4). The multiple cloning site (MCS) was found from 79bp to 127bp and the GFP was found to be from 128bp to 571bp. There was only one reading frame present, which suggests that there was no stop codon present between the Calcineurin PCR product and the GFP sequence present in the p...
... middle of paper ...
... to verify the presence of the insert because the restriction enzyme would uniquely cut (cut in only once place) the plasmid at specific locations and produce two bands. The insert size is 522bp and the plasmid is 6156bp resulting in a total 6678 bp. For verification, the plasmid could be cut using BglII and BstBI and this would result in two bands; the plasmid would be about 5481bp and the insert would be about 1197bp. There would be 4 controls with this: the plasmid without enzymes, the plasmid with insert without enzymes, the plasmid with insert with BglII and the plasmid with insert with BstBI. The samples, when loaded on an agarose gel and through electrophoresis, would result in bands to verify that not only did the enzymes would, but the insert was actually in plasmid and this would be seen when the gel is analyzed through chemiluminescence such as SYBR safe.
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