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Fundamental Genetics
Lecture 9
DNA Structure
and Analysis
John Donnie A. Ramos, Ph.D.
Characteristics of the
Genetic Material
Francis Crick
Replication
Storage of
information
Expression of
information
Variation by
mutation
Transformation
might be due to the
polysaccharide
capsule or some
compound required
for capsule
synthesis
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Hershey-Chase Experiment
DNA (and not protein) is the genetic
material in phage T2.
DNA Structure
DNA Structure
Proposed by Watson
and Crick in 1953
based on:
Base composition
analysis of
hydrolyzed samples
of DNA
X-ray diffraction
studies of DNA
Sequence of
nucleotides codes for
the genetic
information (4n where
n refers to the no. of
nucleotides)
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DNA Structure
Nucleotide Linkage
Chargaff Rule
Amount of A is proportional to T
while C is proportional to G
Sum of purines (A+G) equal to
sum of pyrimidines (C + T)
Percentage of G + C does not
necessarily equal to percentage of
A+T
Types of DNA
B DNA
A DNA
Z DNA
Criteria
10 bp
11 bp
12 bp
34 A
37.4 A
40.8
Diameter of helix
20 A
23 A
18 A
Direction of turn
Right-handed
Right-handed
Left-handed
Present
Modified
Absent
Major groove
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RNA Structure
Ribose sugar
Same nitrogenous bases as DNA except that T replaced by U
Single stranded (but can form double strands)
Forms:
Ribosomal RNA (rRNA)
Messenger RNA (mRNA)
Transfer RNA (tRNA)
Molecular Hybridization
Annealing of nucleic acid
(DNA or RNA) strands
sharing nucleotide
sequence similarity
Used to identify
homologous genes in
different species
Reassociation Kinetics
Measures the rate of annealing between
complementary strands
Measures half reaction time (point when
of the reaction are double stranded)
Half Reaction is lower in smaller genomes
Used to measure repetitive DNA
sequences (characteristic of eukaryotes)
Example: In situ
hybridization or
Fluorescence in situ
hybridization (FISH)
Electrophoresis
Agarose gel
electrophoresis
Polyacrylaminde gel
electrophoresis
Separates nucleic acids
by size under an
electrical field
DNA is negatively
charged (travels to +
charge)
Southern Blot
detection of DNA
Northern Blot
detection of RNA
Genbank
http://www.ncbi.nlm.nih.gov/genbank/
Under Search look for
nucleotide
Enter accession number,
author or key words in
the 2nd search box
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