Imagine that you need pUC57 plasmid specially designed with an insertion of ZMI52 gene inside it (which means it is a pUC57+ZMI2 plasmid). For an experiment, you need to transform 2ng of the plasmid into Saccharomyces cerevisiae cell. But you only have 0.5 ng of the plasmid. Now that you have a timeline to perform this experiment and present the result in a conference. You do not have enough time to order a new plasmid. How can you arrange the required amount of plasmid to perform your experiment? (Note that you cannot take it from a friend working in a lab!) Discuss the pros and cons of your adopted method. Keep in mind, you can propose solutions, but you must think about troubleshooting at the same time. The solution may not work on this plasmid!!
Hi there,
As it is a pUC-based plasmid, why don't you just transform E. coli with it (Ampicilline selection) and make a plasmid miniprep from an overnight culture of an isolated clone??? You would get µg of it!
Are you sure it is actually 2ng you need? not 2µg? 2ng sounds as a very low amount for yeast transformation...
Hi there,
As it is a pUC-based plasmid, why don't you just transform E. coli with it (Ampicilline selection) and make a plasmid miniprep from an overnight culture of an isolated clone??? You would get µg of it!
Are you sure it is actually 2ng you need? not 2µg? 2ng sounds as a very low amount for yeast transformation...
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