Brief Review of DNA Replication:
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Brief Review of DNA Replication:. Nearest Neighbor Analysis (Kornberg 1956):. In vitro DNA synthesis •Template DNA •DNA polymerase •dNTPs. *dATP. *dCTP. *dGTP. *dTTP. •Allow DNA replication to take place •Denature DNA •Treat with micrococcal nuclease-

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Brief review of dna replication

Nearest Neighbor Analysis (Kornberg 1956):

In vitro DNA synthesis

•Template DNA

•DNA polymerase

•dNTPs

*dATP

*dCTP

*dGTP

*dTTP

•Allow DNA replication to take place

•Denature DNA

•Treat with micrococcal nuclease-

yields mononucleotides with 3’ terminal phosphates


Brief review of dna replication

ssDNA treated with micrococcal nuclease:

A

A

C

C

C

*

G

G

T

T

T

T

C

ssDNA:

*


Brief review of dna replication

Exercise:

List all possible dinucleotides (analysis of nearest 5’ neighbor)

Which must be present in equal frequencies if the DNA strands are:

Parallel ?Antiparallel?


Brief review of dna replication

ApA

ApC

ApG

ApT

CpA

CpC

CpG

CpT

GpA

GpC

GpG

GpT

TpA

TpC

TpG

TpT

Exercise:

List all possible dinucleotides (analysis of nearest 5’ neighbor)

Which must be present in equal frequencies if the DNA strands are:

Parallel ?Antiparallel?

ApC = TpG CpA = GpT

ApG = TpC CpG = GpC

ApT = TpA CpT = GpA

ApC = GpT CpA = TpG

ApG = CpT GpA = TpC




Dna melting
DNA Melting

*Temperature at which DNA strands are ½ denatured is the melting temperature or Tm

*GC content of DNA has a significant effect on Tm