MICR 4500 Lecture Notes - Lecture 9: Open Reading Frame, Replica Plating, Dna Supercoil
30 Dec 2019
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Shotgun cloning: whole genome of dna is cut with re or physical breakage. Directional cloning: put an insert in a specific/known direction in a clone w digestion. Use 2 res to maintain the open reading frame w cis acting regulatory elements. Pcr based cloning: make copy of piece of dna and insert into plasmid. Transformation: force cells to take dna from the environment. Electroporation: create a pore in cell to allow dna uptake. Chemical induced: use sodium chloride, cold and heat to make cells take up dna. Selection: look at what mutants are there in general. Screening: requires testing for thousands of colonies for a specific trait. Radiation: lead to point and deletion mutations. Replica screening: pick and patch v velveteen transfer. Conditional screen: any mutation that only affects the cell under certain conditions. Under other conditions the cell is wild type. Functional structural screen: find mutations that prevent things such as taxis and adherence.
