Bio1AL_Fa09_lab5_prelab

Bio1AL_Fa09_lab5_prelab -...

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Unformatted text preview: Plasmid
isolation
and
digestion,
Complementation
II,
and
GMB
I
Pre‐lab.

Due
at
the
start
of
lab.
 
 Name

 
 
 
 
 

GSI
&
Sect
#

 
 
 
 
 
Station
#
 
 
 
 To
answer
these
questions
read
all
of
the
lab
material
on
constructing
recombinant
DNA
(pages
64‐90)
even
 though
you
are
only
doing
pages
77‐84
for
lab
5.

You
also
need
to
read
about
complementation
(pages
111‐118)
 even
though
you
are
only
doing
pages
111‐113.

You
also
need
to
read
about
GMB
I
(pages
135‐144).

Write
 legibly.

You
will
NOT
be
admitted
to
lab
unless
you
turn
in
this
completed
pre‐lab
at
the
start!

 
 1)

Define
the
following
terms:

 
 binding
affinity:
 
 
 crossing
over:
 
 
 homologous
chromosomes:
 
 
 complementation:
 
 
 restriction
enzyme:
 
 
 
 2)


Can
yeast,
deficient
in
histidine
biosynthesis,
grow
on
YPD
media?
on
SD
media?
 
 
 3)

Where
do
you
dispose
of
the
used
pipette
tips
and
used
centrifuge
tubes?
 
 
 4)

Discuss
the
DNA
binding
affinity
of
the
spin
column
and
how
its
affinity
changes
with
changes
in
ionic
 concentration.
 
 
 
 5)

During
the
sequencing
reactions
either
single
stranded
DNA
or
double
stranded
DNA
is
used
as
a
template
(if
 double
stranded
DNA
is
used
it
must
be
denatured
to
yield
single
stranded
DNA).

Primers
are
then
added
which
 are
complementary
to
one
of
the
strands.

The
primers
anneal
and
provide
a
free
3’
OH
group
to
which
 nucleotides
can
add.

Nucleotides
base
pair
with
the
template
strand
and
are
linked
in
a
5'
to
3'
direction
by
DNA
 polymerase.


 a)

In
a
typical
reaction
0.5
µg
of
DNA
is
used
for
the
sequencing
reaction.

If
the
DNA
is
single
stranded
and
is
 4,000
nucleotides
long,
how
many
molecules
of
DNA
is
present
(in
the
0.5
µg
of
DNA)?

The
average
molecular
 weight
of
a
nucleotide
is
300
grams/mole.

Show
your
work
and
include
all
units
(include
cancellation
of
units).

 Give
the
exact
number.
 
 
 
 b)

Four
reactions
are
carried
out,
each
with
a
ratio
of
300
molecules
of
dNTP's
to
1
molecule
of
ddNTP.

What
is
 the
probability
that
termination
will
occur
for
a
given
position
in
the
growing
strand
when
it
is
opposite
a
T
in
 the
template
strand?

In
other
words,
what
percentage
of
the
time
will
a
ddATP
be
incorporated
versus
a
dATP
in
 the
growing
strand?
 
 Pre­lab

continued
on
the
other
side.
 A25
–
Fall
2009
 
 6)

Double
stranded
DNA
will
convert
to
single
stranded
DNA
if
heated
high
enough;
this
temperature
is
known
 as
the
“DNA
melting
point”.

Which
DNA
sequence
would
you
expect
to
have
a
higher
DNA
melting
point,
one
 with
60%
GC
or
one
with
45%
GC?

Explain
your
answer
based
upon
the
number
of
hydrogen
bonds
a
GC
base
 pair
can
form
(versus
a
AT
base
pair).
 
 
 
 7)

Diagram
the
respective
stages
of
the
mitotic
or
meiotic
cell
cycle
of
a
cell
that
has
a
diploid
(2N)
number
of
six
 chromosomes.

Use
different
colored
pens
or
pencils.

Indicate
some
crossing
over
events.

It
is
CRITICAL
YOU
 UNDERSTAND
MEIOSIS
in
order
to
understand
genetics.
 
 MITOSIS
 
 
 prophase
 
 
 
 
 
 
 
 
 
 
 
 
 metaphase

 
 
 
 
 
 
 
 
 
 
 
 
 anaphase
 
 
 
 
 
 
 
 
 
 
 
 
 

 
 
 
 A26
–
Fall
2009
 MEIOSIS
 prophase
I
 metaphase
I
 anaphase
I
 anaphase
II
 ...
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