Bio103L, Expt 3 & 4 (Revised)

Bio 103 Lab
• Lab Experiment 3 & 4

Functions of Proteins
• Proteins main function is to build, maintain and repair all our body tissues.
• Protein can also be used as energy source by body.
• Some proteins have a protective or defensive function.
• Proteins serve as the carrier and transporters of essential substances.
• Types of proteins: Structural proteins & Regulatory proteins.
 Structural proteins i.e., collagen or elastin
 Regulatory proteins i.e., enzymes

Milk Protein
• Milk contains 3.3% total protein.
• Contain all 9 essential amino acids required by humans.
• There are two major categories of milk proteins,
 casein and
 whey proteins.
• Milk proteins are synthesized in the mammary gland.

Principle
• Reaction of Biuret reagent with peptide bonds forming

chelate complex.
• Cu (II) ion containing biuret reagents, interact with a

compound that contains at least four peptide bonds,
resulting in the formation of violet/purple colored complex.
• A color change from blue to violet indicates that proteins

are present.
• The greater the concentration of peptide bonds, the greater

the color intensity.
Reaction of Biuret reagent with
peptide bonds forming chelate
complex.
Principle (cont.)
• If the concentration of peptide bonds is low, it results the color change is from the blue
to pink.
• Single amino acids (no peptide bonds present) and dipeptide (only 1 peptide bond
present) which will give a negative result.
• Peptides with at least two peptide bonds are required for the formation of the chelate
complex.
EXPERIMENT
• Apparatus
• Procedure
 Glassware
1. Add 2 ml of the milk sample to a clean and dry test tube.
 Test tubes
2. Add 2 ml of biuret regent.
 Test tube holder
3. Repeat the steps above with distilled water to prepare a
 Dropper
negative control.
• Reagents 4. Shake well and allow the mixture to stand for 5 minutes.
5. Observe any color change.

 Biuret reagents
 Test sample (liquid milk)
 Distilled water
Experiment 4
Isolation of plant genomic DNA from spinach
• Deoxyribonucleic acid (DNA) is a nucleic acid that contains the genetic instructions for
the development and function of living things.
• The structure of DNA is called a double helix, which looks like a twisted staircase.
• The main role of DNA in the cell is the long term storage of information's.
• There are four different bases:
 adenine (A), cytosine (C), guanine (G), or thymine (T).
• The two strands are held together by hydrogen bonds between the bases, with adenine
forming a base pair with thymine, and cytosine forming a base pair with guanine.
DNA Extraction
• DNA can be isolated from living tissues e.g., spinach, banana, onion.
• DNA isolated from the nucleus, for which cell wall and plasma membrane must be broken
down by mechanical forces, in the presence of salt and detergent.
• Isolation of DNA from plant material poses special challenges, i.e., several plant metabolites
have chemical properties similar to DNA.
• Young and healthy tissues are recommended to use for DNA extraction
 Young tissues contains more cells than older tissue.
 Young tissue of the same weight contains fewer metabolites.

Principle
• The process of isolating nucleic acid is a crude extraction method.
• By using salt and detergent solution causes the cell membrane to break down or lyse and
emulsifies the lipids and proteins of the cell.
• After incubation period, the crude mixture is filtered to get homogenized solution.
• Pouring of this solution into petri dish and glass tube containing 95% ethyl alcohol or
ethanol, causes the precipitation of DNA in time.
• After some moment, it appears as thick, stringy, white or slight greening mass.
• As DNA is not soluble in alcohol, can be isolated as they clamp together and become
visible to the naked eye.
EXPERIMENT
• Materials • Procedure
 Measuring cylinder, beakers Chop and grind sample
 Mortal and pestle
 Light colored dish washing liquid Add liquid with detergent and salt
solution
 Ethanol 95%
 Large spoon for mixing

Incubate at 56°C for 10 minutes
 Petri dishes
 Strainer/ funnel
 Salt Filtration
 Filter cloth or paper
Add 95% ethanol and visualize DNA

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Bio 103 Lab

• Lab Experiment 3 & 4

• Protein can also be used as energy source by body.

• Some proteins have a protective or defensive function.

• Proteins serve as the carrier and transporters of essential substances.

• Types of proteins: Structural proteins & Regulatory proteins.

 Structural proteins i.e., collagen or elastin

 Regulatory proteins i.e., enzymes

• Milk contains 3.3% total protein.

• Contain all 9 essential amino acids required by humans.

• There are two major categories of milk proteins,

• Milk proteins are synthesized in the mammary gland.

• Reaction of Biuret reagent with peptide bonds forming

• Cu (II) ion containing biuret reagents, interact with a

• A color change from blue to violet indicates that proteins

• The greater the concentration of peptide bonds, the greater

5. Observe any color change.

 Test sample (liquid milk)

• There are four different bases:

 adenine (A), cytosine (C), guanine (G), or thymine (T).

 Young tissues contains more cells than older tissue.

 Young tissue of the same weight contains fewer metabolites.

• The process of isolating nucleic acid is a crude extraction method.

 Measuring cylinder, beakers Chop and grind sample

 Mortal and pestle

 Large spoon for mixing

 Filter cloth or paper

Add 95% ethanol and visualize DNA

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