CARA’S NOTES ⋅˚₊‧ ୨୧ ‧₊˚ ⋅ BIO 024: BIOCHEMISTRY ◌ྀི

DVM1-A4 EXPERIMENT 5 AND 6: NUCLEIC ACIDS & PROTEINS ⋆౨ৎ˚⟡˖

✩₊˚. NUCLEIC ACIDS ❀˖°

3. What does the salt do?
● Helps the DNA strands come
ACTIVITY 1: DNA EXTRACTION
together
The DNA molecule is shaped like a
twisted ladder.
4. What does the cold ethanol do?
● To precipitate the DNA
● DNA is composed of building
blocks called nucleotides:
5. Why can’t you use room
○ Deoxyribose sugar
temperature ethanol?
○ Phosphate group
● DNA is not soluble in alcohol
○ And one of four nitrogen
bases – Adenine, Thymine,
6. How will you describe the
Guanine, and Cytosine
appearance of the DNA you
● Phosphate and sugars of adjacent
extracted both from banana fruit
nucleotides link to form a long
and your cheek cells?
polymer
● The DNA will appear white and
● Ratios of A-to-T and G-to-C are
will form a clump made of
constant in all living things.
string-like strands that wrap onto
● James Watson and Francis Crick
the glass rod

RNA is an intermediary between DNA

and protein.
ACTIVITY 2: MUTATIONS
● Cytoplasm Mutations
● Watson and Crick proposed that ● Changes in genetic information
RNA must copy the DNA message ● Mutations generate novel changes
in the nucleus and carry it out to that are starting points of
the cytoplasm, where proteins are evolution, and some are
synthesised responsible for disease
● Could either be beneficial,
ACTIVITY 1 | DNA EXTRACTION harmful, or neutral
QUESTIONS ANSWERED: ● Sourced of mutation:
○ Ultraviolet radiation
1. Why did you mash the banana ○ Chemicals from a variety of
fruit? man-made and natural
● To help release the DNA sources
○ DNA replication itself (not
2. Why do we use liquid soap? always perfect)
● Breaks apart the cellular and
nuclear membranes and it
releases the DNA
CARA’S NOTES ⋅˚₊‧ ୨୧ ‧₊˚ ⋅ BIO 024: BIOCHEMISTRY ◌ྀི
DVM1-A4 EXPERIMENT 5 AND 6: NUCLEIC ACIDS & PROTEINS ⋆౨ৎ˚⟡˖

Types of Mutations
POINT MUTATIONS
● Silent Mutation
A single base pair change in DNA that
doesn't alter the amino acid sequence in
the protein.
● Missense Mutation
A single base pair change in DNA that
leads to a di erent amino acid being
incorporated into the protein.
● Nonsense Mutation
A single base pair change in DNA that
creates a premature stop codon, resulting
in a shortened, non-functional protein.

CHROMOSOMAL MUTATIONS
● Deletion
Loss of a segment of DNA, which can
disrupt genes depending on the size and
location of the deletion.
● Inversion
Reversal of the orientation of a DNA
segment within a chromosome.
● Duplication
A segment of DNA is abnormally copied
one or more times.
● Translocation
Movement of a DNA segment from one
chromosome to another non-homologous
chromosome.
CARA’S NOTES ⋅˚₊‧ ୨୧ ‧₊˚ ⋅ BIO 024: BIOCHEMISTRY ◌ྀི
DVM1-A4 EXPERIMENT 5 AND 6: NUCLEIC ACIDS & PROTEINS ⋆౨ৎ˚⟡˖

✩₊˚. PROTEINS ❀˖° soluble in water while gluten is

insoluble in water
● Thus, gluten can be separated
● Are naturally occurring,
from starch
unbranched polymers in which
● Iodine solution is used to test the
the monomer units are amino
complete removal of starch, which
acids.
involves the formation of
● More specifically, a protein is a
blue-iodo starch complex
peptide in which at least 40 amino
● Refrigeration keeps the protein
acid residues are present
from degrading

˚ ◌༘ GLUTEN ⊹ ˖ QUALITATIVE COLOUR REACTION:

● Is a mixture of two proteins, BIURET TEST
glutenin and gliadin - Rose-pink to violet then purple
● It is also the composite of a - Due to peptide linkages
prolamin and glutelin, which exist, NINHYDRIN TEST
conjoined with starch, in the - Deep blue colour to violet or
endosperm of various purple
grass-related grains - Presence of free amino group
● The prolamin and glutelin from XANTHOPROTEIC TEST
wheat (gliadin, which is - Yellow w/ HNO3 and to orange w/
alcohol-soluble and only glutenin, NaOH
which is soluble in dilute acids or - Nitration of benzene ring of
alkalis) constitute about 80% of the aromatic amino acids (Tyrosine
protein contained in wheat fruit. (Y), Tryptophan (W), and
○ (from Latin gluten, “glue) Phenylalanine
○ Is a protein composite MILLON’S TEST
found in wheat and related - Flesh to red or brick red complex
grains including barley and with mercurous nitrate in HNO3
rye. - Detects the presence of tyrosine,
○ Gluten gives elasticity to an amino acid with a phenolic
dough, helping it rise and group (OH attached to an aromatic
keep its shape and often ring).
gives the final product a HOPKINS-COLE TEST
chewy texture - Reddish violet ring at the junction
w/ Magnesium salts of oxalic acids
Washings remained clear after the iodine - Detects the presence of
test, indicating removal of all starch. tryptophan, an amino acid
● The principle involved in the containing an indole ring.
isolation of gluten is the di erence
in solubility. The starch is partially
CARA’S NOTES ⋅˚₊‧ ୨୧ ‧₊˚ ⋅ BIO 024: BIOCHEMISTRY ◌ྀི
DVM1-A4 EXPERIMENT 5 AND 6: NUCLEIC ACIDS & PROTEINS ⋆౨ৎ˚⟡˖

SAKAGUCHI TEST SULLIVAN OR SULLIVAN CYSTEINE

- Deep red colour w/ a-naphthol REACTION
and sodium hypochlorite or - Red colour w/ strong reducing
bromite agent sodium hydrosulfite
- Detects the presence of arginine, - Cysteine and cystine qualitative
an amino acid containing a estimation
guanidino group. PAULY DIAZO TEST
LEAD ACETATE TEST - Uses diazotized sulfanilic acid,
- When the test is positive for which reacts with specific amino
cysteine or cystine, a black acid side chains under alkaline
precipitate of lead sulphide (PbS) conditions.
forms due to the reaction between - Detects the presence of tyrosine
the sulphide ions (released from and histidine in proteins.
the amino acids under alkaline
conditions) and lead ions from the HYDROLYSIS OF INTACT PROTEINS
lead acetate. - Chemical reaction in which water
- Detects the presence of amino is used to break down bonds of a
acids containing a sul ydryl particular substance
group (SH) or a disulfide bond - Reaction with water/breaking
(S-S). This includes cysteine and using water
cystine, but not methionine. ACID HYDROLYSIS
LIEBERMAN TEST ● Generally only used in the
- Violet colour determination of tryptophan,
- Tryptophan since other amino acids are
MOLISCH TEST degraded
- Violet ring ENZYMATIC HYDROLYSIS
- Glycoprotein (ex. ● Is rarely used to except for the
Gamma-globulin) determination of glutamine and
SODIUM NITROPRUSSIDE TEST asparagine, which are converted
- When the test is positive for a thiol to aspartic and glutamic acids
group, a red colour develops. together with ammonia by acid
- Detects the presence of thiol hydrolysis
groups (SH), primarily from the - A process in which enzymes
amino acid cysteine. enhance bond cleavage in
FOLIN REACTION molecules with the addition of the
- Deep red colour in alkaline elements of water
solution w/ sodium 1,2 - Determination of glutamine and
naphthoquinone-4 sulfonate asparagine which are converted to
- As quantitative estimation of AA aspartic acid and glutamic acid
ENZYMES
- Lipase: Fats
CARA’S NOTES ⋅˚₊‧ ୨୧ ‧₊˚ ⋅ BIO 024: BIOCHEMISTRY ◌ྀི
DVM1-A4 EXPERIMENT 5 AND 6: NUCLEIC ACIDS & PROTEINS ⋆౨ৎ˚⟡˖

- Amylase: Carbohydrates PRECIPITATION:

- Proteinsases: Proteins Precipitants
● By acids
ADVANTAGES & DISADVANTAGES OF ● By salts of heavy metals
ENZYMATIC HYDROLYSIS ● By alcohol
Advantages Organic acids
▪Mild process ● Trichloroacetic acid
▪ Requires less energy ● Phosphomolybdic acid
▪ Produce less by-product ● Phosphotungstic
Disadvantages ● Picric acid
▪ Low reaction rate ● Tannic acids
▪High yield of sugar monomers
▪Usually incomplete TANNIC ACID
● Topically, tannic acid is used for
DENATURATION cold sores and fever blisters,
● Is the partial or complete diaper rash and prickly heat,
disorganisation of a protein’s poison ivy, ingrown toenails, sore
characteristic 3D shape as a result throat, inflamed tonsils, spongy or
of disruption of its secondary, receding gums, acute dermatitis,
tertiary, and quaternary structural and as a styptic.
interactions ● Tannic acid is used orally and
● Causes inactivation of protein topically for bleeding, chronic
activity diarrhoea, dysentery, bloody
● Causes loss of water solubility or urine, painful joints, persistent
renders the protein insoluble at coughs, and cancer.
isoelectric point ● Vaginally, tannic acid is used as a
● The process of altering the douche for leukorrhea.
native/low free energy ● In foods and beverages, tannic
conformation of a protein acid is used as a flavouring agent.
● Peptide bonds remain intact and ● In manufacturing, tannic acid is
the protein usually retains its used in hemorrhoidal ointments
original primary structure and suppositories, for tanning
● Principle: Denatured protein can hides and manufacturing ink, and
return to its native state and to kill dust mites on furniture.
resume its specific biological ● Tannic acid has astringent e ects
activity ● It dehydrates tissue, internally
reducing secretions, and
externally forming a protective
layer of harder, constricted cells. ▪
Tannins show some evidence of
CARA’S NOTES ⋅˚₊‧ ୨୧ ‧₊˚ ⋅ BIO 024: BIOCHEMISTRY ◌ྀི
DVM1-A4 EXPERIMENT 5 AND 6: NUCLEIC ACIDS & PROTEINS ⋆౨ৎ˚⟡˖

antiviral, antimicrobial, CNS result of high concentration of

depressant, and cariostatic e ects neutral salts
● Some evidence suggests that 2. E ect of pH
tannins might cause cancer, but - solubility is minimal at isoelectric
other evidence shows tannins point and increases with
might prevent it ▪ Regular increasing acidity or alkalinity
consumption of herbs with high 3. E ect of organic solvents
tannin concentrations correlates - organic solvents (miscible with
with increased incidence of water), ex. methanol, ethanol, and
esophageal or nasal cancer . acetone.
- decrease dielectric constant
INORGANIC ACIDS: HNO3 OR NITRIC - Increase electrostatic forces
ACIDS between charged particles in
● Used in detecting the presence of solution
proteins in the urine through 4. E ects of Alcohol
Heller’s test - dilute solution: alcohol reduces
● Results: White precipitate turning the solubility of proteins due to an
yellow increase in the electrical forces
between charged particles in
SALTS OF HEAVY METALS solution
● Reaction: proteins from an - High alcohol concentration:
alkaline solution + salts of heavy protein molecules are dehydrated.
metals = precipitation - MORE PRECIPITATION:
● Ex. lead nitrate, silver nitrate, - 95 % alcohol – less
mercuric chloride germicidal
● Metallic salts - used as antiseptic - 70 % alcohol – with greater
and germicides through germicidal e ect
precipitation
● Ex. AgNO3 - used for cauterization
of wounds and ulcers
● Egg white and milks - antidote for
metallic poisoning

FACTORS AFFECTING PROTEIN

SOLUBILITY
1. E ects of neutral salts (NaCl,
MgSO4, (NH4) 2SO4)
- Salting in: increase in solubility w/
low concentration
- Salting out: aka antisolvent
crystallisation, precipitation as a