Week 3 Tuesday Flashcards
What are the toughest fibers found in nature?
spider silk
What are the 3 main ways to deliver DNA?
transformation, conjugation, transduction
What method of transformation is used to maximize efficiency?
electroporation
What are some barriers of a plasmid at different stages (3)?
DNA uptake, Host defense system, selection and maintenance
What is the purpose of selection markers?
ensure cells are maintaining plasmids
What are antibiotic resistance genes?
commonly deployed selection markers
What are some examples of selection markers?
Fluorescent marker, auxotrophic marker, toxin/anti-toxin system, etc
Plasmids must be able to ______.
replicate
Different Ori can lead to different ________.
copy numbers
What are the copy numbers?
how many copies of the plasmid the cell makes during replication
What are some ways to make “competent” cells?
- ensure correct phase of growth (logarithmic phase)
-add positively charged chemicals to neutralize the environment - Add chemicals to
increase permeability
-add reagents to protect the plasmid
What is the phase in which cells are most competent?
logarithmic phase
What is the purpose of adding positively charged chemical to neutralize the environment?
allows negatively charged DNA to be close to the negatively charged membrane
What is the purpose of adding chemical to increase permeability?
aids in pore formation one a heat shock or electric shock is created
What was the purpose of adding salmon sperm?
protects the plasmid by throwing off the defense system (host endonucleases)
