SAFETY precautions

1. Safety begins with the individual’s personal responsibility .

2. Students have a statutory obligation to protect themselves and
others from hazards resulting from their acts or omissions in the
laboratory .
3. Only the students of groups scheduled are permitted to be in the
laboratory. Visitors or intruders should be asked to leave .
4. Smoking, eating and drinking are not permitted on the
laboratory .
5. Outdoor clothing must be left in a cloakroom. Bags must be
placed on side podia provided for the purpose and not allowed to
obstruct gangways or bench tops .
 SAFETY precautions
6. Suitable laboratory coats must be worn in the laboratory and
removed after leaving . Safety spectacles must be worn when
carrying out any procedure where may be slightest risk of eye
injury; gloves of the appropriate type must be worn when necessary.
7. Students are NOT permitted to do any experimental work unless a
supervisor (demonstrator or a member of staff) is present .
8. Glassware and plastic ware that is being used should be labeled
with marker; this avoids confusion and will help the laboratory
staff .
 SAFETY precautions
6. Suitable laboratory coats must be worn in the laboratory and
removed after leaving . Safety spectacles must be worn when
carrying out any procedure where may be slightest risk of eye
injury; gloves of the appropriate type must be worn when necessary.
7. Students are NOT permitted to do any experimental work unless a
supervisor (demonstrator or a member of staff) is present .
8. Glassware and plastic ware that is being used should be labeled
with marker; this avoids confusion and will help the laboratory
staff .
 Regarding substances and procedures hazardous to health.

1. Where a potential hazard exists in a particular practical lesson, this will be

discussed in the talk before the practical and details of safe working methods
will be highlighted in the practical notes .
2. Students must NOT use unfamiliar equipment or procedures .
3. Mouth pipetting is forbidden for acids, bases and hazardous fluids.
4. All hazardous materials are deposited in a fume cupboard .
5. Never use toxic substances without taking the proper precautions and
arranging for safe working. Use volatile solvents in a fume cupboard .
6. Broken glassware must be placed in one of the special bins provided in the
laboratory .
7. Used plastic pipette tips must be placed in the labeled container on the
bench .
 Regarding waste

1. Bench should be left waste-free and tidy at the end of each

practical work – this reduces potential for accidents and spillages
and is of considerable help to the laboratory staff .
2. Laboratory equipment must be left clean after the practical lesson.
The supervisor or the chief technician instructs about the place of
test-tube or flask washing .
 Regarding accidents
1. All accidents and spillages, including any personal injuries
and damage caused to equipment, must be reported as soon
as possible to the supervisor, the chief technician or other
technicians .
2. Concentrated acid or alkali splashed on the skin: a) flood
the splashed surface thoroughly with water and continue until
satisfied that no chemical remains in contact with the skin.
Soap will help to remove chemical which are insoluble in
water; b) remove all contaminated clothing take care not to
contaminate yourself in the process .
 Regarding accidents
3. Splashes in the eye. Eye protection glasses should be worn
for any work where there is a potential hazard but if accident
occurs: a) flood the eye thoroughly but gently with water;
b) seek medical advice for all eye injuries from chemicals .
4. Burns and scalds. Cool affected area by immersing in cold
water. Speed is essential. Continue for at least 5 minutes or
until pain is relieved .
5. Spillages must be cleared up immediately and the area
decontaminated; they must NOT be left as a hazard to others.
 Gene expression

- Gene expression is the process by which information

from a gene is used in the synthesis of a functional gene
product that enables it to produce end products, protein or
non-coding RNA, and ultimately affect a phenotype, as the
final effect.
- These products are often proteins, but in non-protein-
coding genes such as transfer RNA (tRNA) and small
nuclear RNA (snRNA), the product is a functional non-
coding RNA.
Central dogma of molecular biology
 Spectrophotometry and Colorimetry

Definition: It is a method used for identification and

determinations of substances of low concentration or
these difficultly isolated.

Principle: Light can penetrate colored solutions and a

part of it is absorbed or reflected, the other part is
transmitted. The absorbance (A) is directly proportional
to the concentration of the solution
What is light?
- The term light is used to describe radiant energy, with
wavelength visible to human eye. Wavelengths ranging
between about 400 and 750 nm are visible to human
eye.
- Sunlight is a mixture of radiant energy of different
wavelengths that the eye recognizes as white.
 The major components of a simple photometer

Light from the lamp is passed through a filter or monochromator to select the
desired region of light (max), then light passes through an absorption cell
(cuvette) where a portion of light is absorbed depending on the nature and
concentration of the solution. Light, which not absorbed, is transmitted to a
detector which converts light energy to electrical energy that can be registered on
the meter.
Measurement of concentration using Spectrophotometry

If we have a solution of known concentration i.e.

standard (S) and another with unknown concentration
i.e., test or unknown (U), then:
 Measurement of concentration using Spectrophotometry
- The reagent blank: The composition of the reagent blank should be
identical to that of standard or unknown solutions except for the
substance to be measured is replaced by distilled water in blank tube.
- Standard curve: It is done by plotting readings (Absorbance)
against concentrations of standards; this gives us a straight line that
can be used for determining concentration of unknown substance
using its absorbance.
- Standard solutions containing various known concentrations of the
solution to be estimated and scale readings are recorded. Finally, a
reading is made on the unknown solution and its concentration is
determined by a standard curve (calibration curve)
 Estimation of Serum Proteins By Biuret Reaction
Principle:

Plasma proteins:
• Liver is the primary site for synthesis of plasma proteins
1. Albumin
2. Globulins (α-1 globulin, α-2 globulin, β-globulin and γ-globulin).
3. Fibrinogen
 Estimation of Serum Proteins By Biuret Reaction
Interpretation:
- Total plasma protein conc: 70g/L with a range of
63-79g/L (6.3 – 7.9 gm/dl).

- Albumin: 37-53 g/L (3.7 – 5.3 gm/dl) ,

- globulins: 18-36 g/L (1.8 – 3.6 gm/dl) ,

- fibrinogen: 2-4 g/L (0.2 – 0.4 gm/dl)

- Albumin/globulin ratio = 1.2 – 2.5

 Estimation of Serum Proteins By Biuret Reaction
Increase of total protein without altering the albumin-
globulin ratio in:

1. Dehydration.
2. When a person is standing.
3. Vigorous exercise.
4. Using of some anticoagulants
 Albumin

- It is solely synthesized by the liver. It has ahalf-life ofabout

20-25 days, therefore it is a good marker to assess chronic (not
acute) liver damage.

1. An increase in albumin concentrations is usually

due to dehydration.
 Albumin

2. A decrease in plasma albumin concentration occurs with

excessive losses:
a) In urine as in nephrotic syndrome.
b) Into the intestine as in protein-losing enteropathy.
c) Burns.
d) Severe hemorrhage
e) Reduced synthesis of albumin as in liver disease.
f) Malnutrition, malabsorption.
g) Increased catabolism of proteins as in fevers.