In this experiment, we will isolate plasmid DNA from E. coli cells that potentia
ID: 258668 • Letter: I
Question
In this experiment, we will isolate plasmid DNA from E. coli cells that potentially harbor a pJET plasmid with your unknown gene inserted into the MCS. Describe what you would expect to see on an agarose gel under the following conditions after restriction digestion with NdeI and XhoI:
The plasmid does contain your gene of interest flanked by the XhoI and NdeI sites
The plasmid contains your gene of interest and the XhoI site, but the NdeI site has been lost. (Hint: there are two possible results)
The plasmid does not contain your PCR product at all.
Explanation / Answer
The plasmid does contain your gene of interest flanked by the XhoI and NdeI sites.
Two band on agarose gel (one for insert out and other for plasmid).
The plasmid contains your gene of interest and the XhoI site, but the NdeI site has been lost.
One band on the gel (single site so plasmid will be linearized with gene).
The plasmid does not contain your PCR product at all.
one visible band of plasmid and one invisible band of MCS very small may not be visible.
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