Applications of Reproductions & Genetics :
~
1998
Human Genome Project : 3 2 billion
. base
pairs Ethical
& concerns :
-
·
1st time the whole human
genome had been sequenced Confidentiality : who has the right to access this information ?
Found that 1 5% . was made
up of exons What if there wa s a data leak ?
The rest
non-coding DNA Many people would face discrimination from potential employers/
·
:
was
·
Made up of introns health insurance companies
Long stretches of non
coding DNA between
genes If someone's
genome is sequenced they , may
not want to
Humans have just 20' 500
genes In look were expected know if they have a
genetic susceptibility to
developing a
There were more repetitive DNA sequences than disease. However, this affect close relatives who could
many may
expected in the DNA also this allele So-do they have right to know ?
non-coding carry .
a
·
STR :
short tandem repeat Parents wish to have their child screened
may
·
LTR :
Long tandem repeat
·
Do we tell the child ?
·
Do the parents own information ?
For couples with histories of their
a
family a disease
,
embryos can be screened during IVF
However, concerns over
selecting for other desirable
traits need to be addressed
OOK
genome project
-
The of the of 100 1 000 VHS patients & their relatives
sequencing genome
By comparing the genomes of people who develop particular conditions with
-
those who don't alleles ,
associated with those disorders can be identified
The function of the healthy/diseased alleles can then be investigated
-
, -
A technique used to amplify
Polymerase Chain
a specific piece of DNA
Reaction
·
DNA
polymerase cannot
begin a new chain on its own .
-
It can create billions of copies in
just 2-3 hours It can
only extend an
existing chain Iprovided by a primer
Add to reaction mixture 1) Template DNA (to copied) DNA polymerase extend existing I' direction
be can
only
.
:
an
(cannot extend 35)
2) DNA polymerase
3) Free
template
S
strand
nucleotides x 3
3 .... g
5
↓Strands by DNA Pol
3
4) Primers
.
11 1 11
3 5
PCR
°
:
After 30-35 cycles , 95 C -
Denaturing temperature -
breaks
Hydrogen
-
Extension temperature -
72 % there are billions of copies bonds within template DNA , creating single
-
Optimum temperature for the of the target Sequence strands
thermostable DNA polymerase to
55 % Annealing temperature
-
extend the primers & replicate the
The primers bind to
target sequence
complementary sequences
within target DNA
RESTRICTION ENDONUCLEASES
A
Gel Electrophoresis : ·
These enzymes cut the DNA backbone /both strands
-
tequnique that separates DNA fragments according to length within the DNA
DNA will terminal
is-vely charged so move towards the the Each enzyme of restriction
·
-
,
enzyme recognises a
in an electric field Specific base sequence called a restriction site
It will cut the DNA there .
lane 1 lane 2. . .
- a band
Restriction zones are palindromes -
they read the same forwards
& backwards
e .
.
g EcoR1 (from E . Coli)
-
EcoR1 cuts here
↳
I
X
V
EcoR1 creates a
staggered but with
1 + +
C unpaired bases left on both strands .
"
These are called
sticky ends .
~
1998
Human Genome Project : 3 2 billion
. base
pairs Ethical
& concerns :
-
·
1st time the whole human
genome had been sequenced Confidentiality : who has the right to access this information ?
Found that 1 5% . was made
up of exons What if there wa s a data leak ?
The rest
non-coding DNA Many people would face discrimination from potential employers/
·
:
was
·
Made up of introns health insurance companies
Long stretches of non
coding DNA between
genes If someone's
genome is sequenced they , may
not want to
Humans have just 20' 500
genes In look were expected know if they have a
genetic susceptibility to
developing a
There were more repetitive DNA sequences than disease. However, this affect close relatives who could
many may
expected in the DNA also this allele So-do they have right to know ?
non-coding carry .
a
·
STR :
short tandem repeat Parents wish to have their child screened
may
·
LTR :
Long tandem repeat
·
Do we tell the child ?
·
Do the parents own information ?
For couples with histories of their
a
family a disease
,
embryos can be screened during IVF
However, concerns over
selecting for other desirable
traits need to be addressed
OOK
genome project
-
The of the of 100 1 000 VHS patients & their relatives
sequencing genome
By comparing the genomes of people who develop particular conditions with
-
those who don't alleles ,
associated with those disorders can be identified
The function of the healthy/diseased alleles can then be investigated
-
, -
A technique used to amplify
Polymerase Chain
a specific piece of DNA
Reaction
·
DNA
polymerase cannot
begin a new chain on its own .
-
It can create billions of copies in
just 2-3 hours It can
only extend an
existing chain Iprovided by a primer
Add to reaction mixture 1) Template DNA (to copied) DNA polymerase extend existing I' direction
be can
only
.
:
an
(cannot extend 35)
2) DNA polymerase
3) Free
template
S
strand
nucleotides x 3
3 .... g
5
↓Strands by DNA Pol
3
4) Primers
.
11 1 11
3 5
PCR
°
:
After 30-35 cycles , 95 C -
Denaturing temperature -
breaks
Hydrogen
-
Extension temperature -
72 % there are billions of copies bonds within template DNA , creating single
-
Optimum temperature for the of the target Sequence strands
thermostable DNA polymerase to
55 % Annealing temperature
-
extend the primers & replicate the
The primers bind to
target sequence
complementary sequences
within target DNA
RESTRICTION ENDONUCLEASES
A
Gel Electrophoresis : ·
These enzymes cut the DNA backbone /both strands
-
tequnique that separates DNA fragments according to length within the DNA
DNA will terminal
is-vely charged so move towards the the Each enzyme of restriction
·
-
,
enzyme recognises a
in an electric field Specific base sequence called a restriction site
It will cut the DNA there .
lane 1 lane 2. . .
- a band
Restriction zones are palindromes -
they read the same forwards
& backwards
e .
.
g EcoR1 (from E . Coli)
-
EcoR1 cuts here
↳
I
X
V
EcoR1 creates a
staggered but with
1 + +
C unpaired bases left on both strands .
"
These are called
sticky ends .
