1.

Endonuclease are enzymes that cleave the phosphodiester bond within a polynucleotide
chain e.g. DNA 1 which cut DNA relatively non-specifically.
Exonuclease are enzymes that cleave nucleotides one by one form the end of a
polynucleotide chain, the act as a hydrolysing reaction that breaks phosphodiester bond
at either 3’ or 5’ end.

2. Site-specific endonucleases are enzymes that cleave at specific phosphodiester bonds

within a polypeptide chain usually at 4-8 base pairs in length, whereas the endonuclease
cleaves the phosphodiester bonds anywhere within the polynucleotide chain.
If each enzyme was used to cut the same DNA then the endonuclease will have more
lanes and more bands in each lane compared to the site-specific endonucleases, since
the site-specific endonucleases only cleaves at specific sites then the bands will on be
found at certain locations and in certain lanes in which the DNA contains the sites being
cleaved.

3. Concentration of λ DNA is 0.3 µg/µL, since 3 µL of the DNA was used then:
0.3 µg: µL
x : 3 µL
x = 0.9 µg of DNA was used per solution

4.

The first lane is a standard DNA holder and standard DNA length then sample 1, then sample
2, sample 3, sample 4, sample 5, sample 6. The samples from underneath are cut into (bp) base
pairs starting at 250 base pair until 10 (kbp) kilo base pair.
 5.

Number of DNA Appearance of gel lane &

Enzyme λ Sizes
fragments Explanation

- long and not missing any strands, this

is due to the fact that the is no
No Enzyme 48502 base pairs 0.3 µg/µL restriction enzyme and thus the DNA
is not cleaved and it is long
throughout and continuous.
+ The DNA is slightly shorter than the
standard one. The restriction enzyme
0.5 µg/lane cleaves of and creates only 29
Psti 29
0.4 µg/µL segments of the DNA, since psti
restriction enzyme is specific it
makes the lane look like intervals.
* The lane is almost empty, the
enzyme takes long to cut the DNA
and it is easily overloaded and thus
Scai 5 5µ/µg
cannot cleave the DNA in many
fragments but rather in huge fewer
fragments.

6.
# of cuts on λ
Recognition
Enzyme Source/origin DNA POINT of symmetry
sequences
Type 2 restriction
enzyme, extracted 5’-CTGCA/G-3’
from Providencia 5’-CTGCA/G-3’ symmetry
PstI
stuartii, starts 3’-G/ACGTC-5’
cleavage at 5’- 3’-G/ACGTC-5’
CTGCA/G-3’
Type 1 restriction
5’-AGT/ACT-3’ 5’-AGT/ACT-3’
enzyme, starts to
ScaI 3’-TCA/TGA-5’ symmetry
cleave at 5’-
3’-TCA/TGA-5’
AGT/ACT-3’
 MICROBIAL GENETICS
Experiment 5
RESTRICTION ENZYMES
__________________________________________________

MATEUS S.E
201705714
GROUP: FRIDAY MORNING
Date: 19-04-2019