PCR/Gene Tech/DNA Replication Flashcards
What kind of fragments will restriction enzymes create
Sticky ends/blunt ends
What are the places that restriction enzymes bind to
Restriction sites
What are restriction endonuclease used as and how
Defensive mechanism to protect against viruses
Cuts viruses genetic material into fragments preventing replication
Where are restriction endonuclease found
Bacteria
What does SSB proteins do in DNA replication
Bind to single stranded DNA to prevent them from coiling back onto itself, reannealing of separated strands, and protect degradation by enzyme
What are the uses of PCR
Genetic fingerprinting, recombinant DNA, detecting genetic disease
What happens in the 3rd stage of PCR
Mixture is heated to 72 degrees to allow DNA polymerase to work
Free nucleotides line up with template strand
What happens in the 2nd stage of PCR
Mixture is heated to 55 degrees to allow primer to anneal to strands
Where is the tack polymerase from
Bacteria from hot environments
What does the mixture for PCR contain
Primer, free DNA nucleotides, tack polymerase, DNA sample
What type of amplification is PCR
In vitro amplification
What does PCR do
Amplify a sample of DNA to create a large sample for analysis
What happens in the 1st stage of PCR
Mixture is heated to 95 degrees to break H bonds
What are the 3 main step of PCR
Denaturing, Annealing, Extension
