Week 2 Wednesday Flashcards
Is E. coli a prokaryote or eukaryote?
prokaryote
What is E. coli also referred to as?
model organism
What is E. coli?
An opportunistic pathogen, also a prokaryote
E.coli is an industrial workhorse for ___ and _______.
protein and enzyme production
Compare and Contrast E. Coli vs S. cerevisiae.
E. coli is a prokaryote with few organelles and a 20-minute doubling time. S. cerevisiae is a eukaryote that have more organelles and a 100-minute doubling time.
Explain installing hardware in E. coli.
new ‘software’ is easier to ‘install’ although hardware limitations prevent more complex applications
What kind of applications is E.coli good for?
simple plasmid propagations
What makes E.coli good for simple plasmid propagation?
fast growth and well-characterized parts
What is the standard process of plasmid production and delivery?
DNA assembly, propagation in E. Coli, plasmid extraction, and delivery to host
What is needed since the plasmids need to replicate in two different hosts?
two different sets of plasmid parts
What is the purpose of alkaline lysis in plasmid prep?
break down cell wall, denature protein and RNA, converts double-stranded DNA to single-stranded DNA
What are the three steps of plasmid prep?
alkaline lysis, neutralize, clean and concentrate
What is the purpose of the neutralize step in plasmid prep?
convert single-stranded DNA to double-stranded DNA, precipitate single-stranded DNA and proteins
What is the purpose of the clean and concentrate step in plasmid prep?
remove salts, buffers, and residual impurities
How is DNA concentration usually measured?
by detecting bound fluorescent dyes (fluorometer) or measuring the absorbance of UV light
