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DNA Replication and Manipulation (DNA Replication (Coordination of leading…
DNA Replication and Manipulation
DNA Replication
model predictions
semiconservative model
conservative model
dispersive model
new DNA strands grow
replication fork
DNA always grows in 5' --> 3' direction
DNA polymerization: 3' OH at growing end of polynucleotide chain attacks triphosphate group at 5' end of incoming nucleotide (nucleotide triphosphate)
DNA poly --> releases 2 phosphates
Leading versus Lagging
leading
synthesized in one continuous strand
lagging
made in okazaki fragments
DNA poly III removes RNA fragments and adds DNA fragments
primer made by RNA polymerase (RNA primase)
Coordination of leading and lagging strand synthesis
Preparing DNA for copying
helicase
topoisomerase
single-strand binding protein
Copying DNA
primase
DNA poly III
Finishing up
DNA poly I
DNA ligase
Once okazaki fragments are reached, loops around
Proofreading
DNA cleavage function of DNA poly
exonuclease activity
Replication of Chromosomes
Start of replication
eukaryotes = many origins of replications
Telomeres
Isolation, Identification, and Sequencing of DNA Fragments
PCR
Electrophoresis
Restriction Enzymes
Genetic Engineering