Physical Mapping of DNA

Balcera, Katrina Marie

Hepiga, Adah Mae
Overview
Background
Introduction to Physical Mapping
Restriction Mapping Figures
Background

Given a sequence of DNA, how do we figure out

where on some larger chromosome the sequence
lies?

?
Background II
Look for markers that match in both the
chromosome and the shorter sequence.
Markers: Usually short, precisely defined sequences

match!
Background III
How do we create the original map?

Generate fingerprints (markers) with:

Restriction site mapping
Overview
Background
Introduction to Physical Mapping
Restriction Mapping Figures
INTRODUCTION
is a map of known restriction sites within a sequence of
DNA. Restriction mapping requires the use
of restriction enzymes.
A map based on such physical characteristics is called,
naturally enough, a physical map.
INTRODUCTION II
Restriction sites:
A restriction enzyme recognizes a specific short sequence of
nucleotides, called restriction sites and cuts a DNA molecule
at any position that displays the specific sequence.
For example, EcoRI recognizes the sequence: 5 G
A A T T C 3 3 C T T A A G 5 in a DNA molecule and
cuts it after the G : 5 G A A T T C 3 3 C T T A A G
5
INTRODUCTION III
Physical map consists of a list of locations of markers along
the chromosomes of the genome. Such a marker is usually a
known DNA sequence that occurs somewhere in the
genome.
For example, assume we are given a physical map A, B, C,
D, E of a genome G. Let S be a fragment of G with markers
B, C, then we can place S in G: B C A B C D E
Some of the most commonly used
restriction enzymes:
Overview
Background
Introduction to Physical Mapping
Restriction Mapping Figures
Southern Blot Procedure:
DNA is cut into pieces by restriction
enzymes
DNA fragments are separated by gel
electrophoresis
DNA is denatured (by alkali) to produce
single-stranded bands of DNA and
transferred from gel to hybridization filter
Physical Mapping
A physical map is primarily based on the locations
of landmarks along a DNA molecule and units of
distance are expressed in base pairs.
Is constructed by cutting a DNA molecule into man
short fragments and arranging them in order by
identifying overlaps.
A physical map can be made by isolating DNA
from a chromosome then cutting it using restriction
enzyme( also known as restriction endonucleases
)to construct a pattern.