Following is my journal documentation during Master's in Biotechnology completed in 2015. I do understand many changes would've occurred in the curriculum since then, but the basics seldom change. Kindly absorb as per your need.

1. Journal lIndex
PSBTP101:Biochemistry
Ex. No. Staff Member's
Signature
Experiment Title No. Date
Preparation of buffers used in laboratory
(Phosphate, Citrate, acetate and Tris buffer)
1
2 Detection of LDH isozymes by
electrophoresis. 30813
3 Protein estimation by Bradford's method
10 2/813
Study of protein complexes using PAGE and
13
detection with CBB and Silver staining. 26 813
PSBTP102:Immunochemistry P. No. Date Staff Member's
Experiment Title
Signature
Ex. No.
1. Perform serum electrophoresis (horizontal)
24 5/818
Perform iso-agglutination titre
2 24/F 18
2.
Perform Affinity chromatography for
purification of antibodies from serum.
| 3. 29
Perform SDS and native PAGE for studying 33 |2/e|s rN
4.
immunoglobulins structure
PSBTP103: Genomes and Transcriptomes
Staff Member's
Signature
P. No. Date
Ex. No.
Experiment Title
37
39 |o1818
42 113 ye
1.
Restriction digestion reactioon
2. Demonstration of ligation reaction.
3. Perform transformation of bacteria
PSBTP104: Biophysics
Ex.No.
P. No. Date Staff Member's
Experiment Title
Signature
Viscosity studies of proteins (Std BSA & varying
concentration of urea.) 2/71a
1. 4+

2. i

3. M.Sc. I
Biophysics 47
EXPERIMENT NO: 3
Viscosity studies of proteins (Std BSA & varying concentration of urea)
Aim
To determine the changes in the conformation of Albumin by viscosy
measurement
Principle
Protein Conformation is the characteristic 3-dimensional shape of a
protein, including the secondary, super se condary (motifs), tertiary (domains) and
quaternary structure of the peptide chain. Protein structure, quaternary describes
the conformation assumed by multimeric proteins (aggregates of more than one
polypeptide chain).
Denaturation of proteins involves the disruption and possible destruction of
both the secondary and tertiary structures. Since denaturation reactions are not
strong enough to break the peptide bonds, the primary structure (sequence of
amino acids) remains the same after a denaturation process. Denaturation
disrupts the normal alpha-helix and beta sheets in a protein and uncoils it into a
random shape.
Denaturation occurs because the bonding interactions responsible for the
secondary structure (hydrogen bonds to amides) and tertiary structure are
disrupted. In tertiary structure there are four types of bonding interactions
between "side chains" including: hydrogen bonding, salt bridges, disulfide bonds,
and non-polar hydrophobic interactions. which may be disrupted.
Therefore, a variety of reagents and conditions can cause denaturation.
The most common observation in the denaturation process is the precipitation or
coagulation of the protein. High concentration of urea causes denaturation of
proteins and loss of conformation by unfolding due to weakening of bonds that
maintain the tertiary structure. This causes the molecule to be less compact,
leading to increase in the viscosity of the solution compared to that of native
protein molecule.
Viscosity, in general is the resistance of a liquid to its flow. Resistance is
offered when one part of a fluid is moved past another. The force required to slip
one layer of a fluid past another with a given viscosity is called shearing stress
while the rate of movement is called rate of shear. Resistance to this movement is
called the viscosity. The viscosity of a liquid can therefore be defined as the force
per unit area necessary to maintain a unit velocity between two parallel planes of
the liquid separated by a unit distance.
The Ostwald Viscometer basically consists of glass tube in the
shape of U held vertically in a controlled temperature bath. In one arm of U is a
vertical section of precise narrow bore (the capillary). Above this is a bulb, there
Department ofBiotechnology, RD. NationalandWA. Science College, Mumbai

4. (o)
B
B
Key
a An atual 0stuwalds Visconneten shauing
A and B ab the two mahks
b
Diageanntndic hephesentahion of a
Viscamden

5. Cond o SA and Conc of 85A Unea
aph
in6t dine in bctonals SCALE
On Y-axis: Icm 0-25M
On Y-axis: lom 5Les
Lonc o UAca Cond o Uhca and BSA

6. Obsewation lable
Qel AmOunto
S Conc o
Uhea(gms)
(see)(n/no)
0-4841
No Uhea (M)(ec) (se)
3
1-18
0-5 93
6
01914-
I19
g0-3 95 3
0:184 2
I-18
q0 06
18
17 0 44 81
6 112
-18 0-424+ 24
q8 t16
Amount o 6SA in al (on cetrations = 0-1gms
ColculaticnS
x
do
Po
Foamula
TLo
Whene P= Deneity o Uheo-32 gms/cm
Po ensity o K 484 gmscm
:32 = 0 665
Po 924
foh 0-5 M onc of Uhea,
0-665 x 1-18 4do-18)
0-84
b) Fox 1M Cond of Uhea,
0-665 X 19
UAea,
(4,4ol-19)
el
0 4914
fon 2M Con o Uhea,
foh
0-665x :19 (dHo1e)
4844

7. Groph o Cond of DAea agains
SCALE
On
X-axis: lcm= 0 025M
On Y- axis: ICm 0-00Spoise
Aet (nno)
ConC o Unca (m)-

8. M.Sc. I
Biophysics 48
is another bulb lower down in the other arm. In use, liquid is drawn into the upper
bulb by suction and then allowed to flow down through the capillary into the lower
bulb. Two marks (one above and one below the upper bulb) indicate a
known
volume. The time taken for the level of the liquid to pass between these marks is
proportional to the kinematic viscosity.
Formula:
Poiseville equation of viscosity:
(gr4t T)
8 1.v
Where,
= viscosity
= density
g gravity
rE radius ofcapillary
t= time
l= length of capillary
V = volume of liquid
Formula used for practical purpose:
hl ho=( | o) (t/to)
Where,
=
viscosity ofsample protein
=
density of sample protein
t= time (sample protein)
o= viscosity of reference solution
o density of reference solution
to time (reference solution)
The unit of viscosity is poise. The official S.l. unit is Pascal-second.
Requirements:
Sample:
Protein sample
Glass wares
10 ml pipette
1ml pipette
Reagents-
potassium
solution- 100mM/litre
INSTRUMENTS-
chloride (KCI)
Ostwald viscometer
urea solution (0.5M, 1M, 2M,
3M, 4M, 6M, in 100mM/l KCI)
Preparation of solutions:
PREPARATION OF 10OmM KCI
Molecular wt: 74.56
Department ofBiotechnology, RD. Nationaland WA. Science Colege, Mumbai

9. 49
M.Sc. I
Biophysics
74.56 g KCI in 100Oml
0.7456 g KCIl in 100 ml =100mM
Hence 3.728 g KCl in 500 ml = 100mM
1M
=
PREPARATION OF Urea solution
Molecular wt: 60
60 g of urea in 1000 ml 100mM KCI solution = 1 M
1.5 g ofurea in 50 ml 100mM KCI solution 0.5 M
3 gofurea in 50 ml 100mM KCI solution =1 M
6 g of urea in 50 ml 100mM KCI solution = 2 M
9g of urea in 50 ml 100mM KCI solution = 3 M
12 g of urea in 50 ml 100mM KCI solution = 4 M
18 g of urea in 50 ml 100mM KCI solution = 6 M
(To each of the above solution add 1ml of albumin taken from egg)
PROCEDURE:
1. Rinse the viscometer with KCL solution and place it in vertical position in
water bath by carefully clamping one limb
2. Check that it is vertical using plumbline and introduced exactly 20mL(or the
volume marked on the viscometer )of KCI solution at 30c into the bulb A with the
syringe or pipette.
3. Leave for five minutes to equilibrate ,
then either apply positive pressure to the
wide limb(l) or gentle suction to the other limb(1I)until the meniscus rises above
the upper graduation mark B
4. Release the pressure and measure the time(to the nearest 0.1 s) for the liquid
to flow between the two graduation marks B and C
5. Repeat the experiment until the flow times agree within 0.2s and calculate the
average flow timne
6. Repeat the whole procedure with the urea solution alone(to), which are thne
solvent, and then with the bovine serum albumin dissolved in the urea(t1)
7. Plot the values of to ti1and against the concentration of urea and join up the
points with smooth curvesS.
8. Select convenient concentration of urea and calculate the relative viscosities
(t1/to) using the values from the curves.
9. Finally, prepare a graph of the relative viscosity against the concentration of
urea.
RESULT: The viscosity feplein (8sA) intheases wth inchease in
(onstant teropelatine (30'c)
Con certtrabian of UA ea
cONCLUSION:
Department of Biotechnology, RD. National and WA. Science College, Mumbai

10. d) fo 3M conc o ea ,
Lhet 0-665x I-17 ( |do = 14)
0-7121
fo 4M (Onc oDAea
Lel 0 665 x 18 t t H8)
0-+347
Con clubian
Using Otuwald s Viscometen, the vislosiby O Polein
BSA) was found to inu.ease with
UARa his is
inacase in Concemration
do denatuatian Peotein by Uea ie
clue
he eakening nydhaphobic bands PLoteins Jeading to a
un
change p.eotein ContoAmcction AeSuting in a ess Co0p act
un
vis (osity
peoten moleule
Thu, it
wth a highe
can be tnfused that UAea can be ed a a
denahuinq aqent