Recognition sequences
Restriction enzyme
EcoRI cuts the DNA
into fragments.
Sticky
end
• Originally protected bacteria
from viruses.
•Restriction Site = Recognition
sequence NOT Cleavage site
•Recognition site is a planidrome
(just like
“radar”
“Madam, I’m adam”
Example of
Restriction
Sites:
Palindromes:
Journal 12.09.10:
1. Which sizes of DNA fragments
do you expect to get from the
digestion?
2. How can one use the
electrophoresis procedure to
estimate the size of the DNA
fragments?
EcoRI HinDIII BamH1
2300
4000
3000
5000
1. Which fragment sizes will
result from cutting with:
A. HinDIII alone? B. EcoE1
alone?
C. Both HinDIII+EcoR1
2. Draw the sizes into the gel.
bp
2300
E
EcoRI
HinDIII
BamH1
H
B
4000 3000 5000
bp
23000 bp
9000 bp
6000 bp
4000 bp
2300bp
2000 bp
500 bp
To Students that are presenting
about restriction maps and
expected sizes:
You may want to peak through
the slides about gel
electrophoresis, to see the
connection between your part
and the next – electrophoresis.
Restriction digest of pARA-R
Recombinant plasmid of interest
pARA-R
4720 bp
rfp
702bp
pARA-R construct
Recombinant plasmid of interest
pARA-R
pARA-R
4720 bp
4720 bp
rfp
rfp 702bp
702bp
Expected fragments
pARA-R construct
Recombinant plasmid of interest
4720-702=4018bp
pARA-R
pARA-R
4720 bp
702 bp
4720 bp
rfp
rfp 702bp
702bp
restriction analysis of pARA-R
Restriction fragments after digest with Hind III and BamH I
BamH I
Hind III
4018 bp
BamH I
Hind III
702bp
Additional Resources:
Restriction Map Exercise for
high-school (if you want to use
this)