As the size of the protein will be larger, more difficult it will be cloned. Can u guys suggest a good way to get the desired mutations in my tagged gene ? Waiting for the answer.
Dear Rudrakshi Sharma
what do you mean for the size of the tagged gene is bigger?
How much is big the entire vector that contain the gene?
Generally i'm using the PIPE cloning approach (see attached papers) to perform insertion/deletion/mutagenesis in my existing clones.
It work quite well if the size of the plasmid is not too big (
Hi there,
Important issue : is it the gene located on a plasmid or is it genomic?
I don't understand why the mutatded gene would be bigger apart from the case of frameshift mutation which sometimes results in a bigger gene product... Can you explain more precisely this particular point please? Thanks
The plasmid which I am using is a CEN plasmid for cloning but in comparison to it my desired mutated gene product which will be ligated to it is very large . And if its tagged , it becomes more difficult to be ligated. So I need help with this. How can I clone my mutated gene to the plasmid ? Thanks for the previous answers.
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