Hi, I really need help doing a molecular cloning experiment.
I am a Ph.D student doing a rotation in a lab that does molecular cloning all the time but I am super confused with all the explanations I am getting.
Basically, I need to make a DNA construct consisting of 3 different DNA sequences.
2 of the DNA sequences we need to isolate from a mice tissue and the third DNA seq. is in a vector.
Then, I will do the Gibson-Assembly PCR experiment to put that those 3 inserts into a plasmid.
Now, my confusion is in the primer designing part. Do we need to design the primers to isolate the DNA sequences from the tissue or are the primers used in the Gibson-Assembly run?
Another question is, how does one isolate those DNA sequences?
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The Science Buddies Ask the Experts forum is for students in grades K-12 who are doing hands-on science projects. There are other sites on the web where your questions would be more appropriate and likely to get useful responses.