To understand all the equipment and to know about the principle and their uses.

1. SUBMITTED BY:
Likhit Patnaik
Regd.no- 201221211
M.Sc. Ag
Department of Agricultural
Bio-technology
TOPIC : Equipment required for Biotechnological lab & its Application

2. Objective of Laboratory
1. To Identify common lab equipment pieces and to describe
the principle and their function.
2. SOPs should be followed specifically while in the
laboratory.
3. Understand the role of equipment and chemical reagents
used while doing the experiment in a laboratory.

3. Application of Biotechnology
1. Industrial biotechnology
2. Diagnostics
3. Genetically modified crops for agriculture
4. Processed food
5. Bioremediation
6. Biotechnology in medicine
7. Safety Concerns

4. Biotechnological laboratory Equipment
• pH-Meter: It is an electronic instrument used for measuring
the pH of a liquid (acidity or alkalinity).
• A pH meter consists of special measuring probes connected
to an electronic meter that measures and displays the pH
reading.
• pH meter was invented by Arnold Orville Beckman in 1936.
• Principle: It defined as the –ve logarithm of hydrogen ion
concentration.
• Types: Manual pH meter and Digital pH meter
• Probe types: 1. Glass electrode
2. Reference electrode
3. Combination gel electrode
Manual pH meter
Digital pH meter

5. Glass electrode Reference electrode Combination gel electrode

6. Ec-Meter: It is used to measures the electrical conductivity in a
solution.
• It is commonly used in hydroponics, aquaculture, aquaponics,
and freshwater systems to monitor the amount of nutrients,
salts or impurities in the water.
• It is highly depend on temperature, reciprocal to resistivity.
• The SI unit of electrical conductance is defined as Siemens per
meter (S/m), and denoted by the symbol “σ.”
Ec meter
Working Principle: A conductivity meter encompasses four electrodes and leverages
potentiometric technique to measure the rate of conductivity. The electrodes are manufactured
using platinum material, cylindrical structured, and placed concentrically. The outer pair of
electrodes is subjected to externally applied AC current and potential difference across the
inner material is calculated using ohm law along with other parameter considerations such as
distance amongst electrodes and surface area.

7. ANALYTICAL CENTRIFUSE
INVENTED BY: Theodor Svedberg
A centrifuge is a equipment that puts an object in rotation
around a fixed axis applying a potentially strong force
perpendicular to the axis of spin (outward).
PRINCIPLE: The centrifuge works using the sedimentation
principle, where the centripetal acceleration causes denser substances
and particles to move outward in the radial direction. At the same
time, objects that are less dense are displaced and move to the center.
USES: centrifuge can spin at up to 15,000 rpm to facilitate separation of the different phases of the
extraction. In DNA extraction To move precipitated DNA to the bottom of the container and make it
stick there, so that the supernatant can be poured off without losing your extract. To separate cell
debris from DNA-containing supernatant, so that this supernatant can be removed and DNA can be
precipitated out of it.

8. THERMAL CYCLER
- Developed by Kary Mullis
-The thermal cycler (also known as a thermocycler, PCR machine or
DNA amplifier) is commonly used to amplify segments of DNA via the
polymerase chain reaction (PCR).
Working principle of PCR. As the name implies, it is a chain reaction, a small fragment of the
DNA section of interest needs to be identified which serves as the template for producing the
primers that initiate the reaction. One DNA molecule is used to produce two copies, then four, then
eight and so forth.
There are three major steps in a PCR, which are repeated for 30 or 40 cycles. This is done on an
automated cycler, which can heat and cool the tubes with the reaction mixture in a very short time.
Denaturation at 94°C:
During the denaturation, the double strand melts open to single stranded DNA, all enzymatic
reactions stop (for example: the extension from a previous cycle).

9. Annealing at 54°C:
The primers are jiggling around, caused by the Brownian motion. Ionic bonds are constantly
formed and broken between the single stranded primer and the single stranded template. The more
stable bonds last a little bit longer (primers that fit exactly) and on that little piece of double
stranded DNA (template and primer), the polymerase can attach and starts copying the template.
Once there are a few bases built in, the ionic bond is so strong between the template and the primer,
that it does not break anymore.
Extension at 72°C:
This is the ideal working temperature for the polymerase. The primers, where there are a few bases
built in, already have a stronger ionic attraction to the template than the forces breaking these
attractions. Primers that are on positions with no exact match, get loose again (because of the higher
temperature) and don't give an extension of the fragment.
The bases (complementary to the template) are coupled to the primer on the 3' side (the polymerase
adds dNTP's from 5' to 3', reading the template from 3' to 5' side, bases are added complementary to
the template).

10. GEL ELECTROPHORESIS CHAMBER
Invented by: Arne Tiselius in the 1931.
Gel electrophoresis chamber is an equipment for separation and analysis
of macromolecules (DNA, RNA and proteins) and their fragments,
based on their size and charge.
Principle:
"Electrophoresis" refers to the electromotive force (EMF) that is used to move the molecules through the gel
matrix. By placing the molecules in wells in the gel and applying an electric field, the molecules will move
through the matrix at different rates, determined largely by their mass when the charge to mass ratio (Z) of all
species is uniform. However, when charges are not all uniform then, the electrical field generated by the
electrophoresis procedure will affect the species that have different charges and therefore will attract the species
according to their charges being the opposite. Species that are positively charged will migrate towards the
cathode which is negatively charged (because this is an electrolytic rather than galvanic cell). If the species are
negatively charged they will migrate towards the positively charged anode. Nucleic acid molecules are separated
by applying an electric field to move the negatively charged molecules through a matrix of agarose or other
substances. Shorter molecules move faster and migrate farther than longer ones because shorter molecules
migrate more easily through the pores of the gel. This phenomenon is called sieving. Proteins are separated by
charge in agarose because the pores of the gel are too large to sieve proteins.

11. Uses:
It is used in biochemistry and molecular biology to separate a
mixed population of DNA and RNA fragments by length, to
estimate the size of DNA and RNA fragments or to separate
proteins by charge.
• Gels used are
-Agarose gel: used for seprating DNA fragments of usually 50-
20,000 bp in size
- polyacrylamide gel :Polyacrylamide gels are usually used for
proteins and small fragments of DNA (5- 500 bp)
- Starch :Partially hydrolysed potato starch makes for another non-
toxic medium for protein electrophoresis.
Types-
1. Horizontal gel electrophoresis
2. Vertical gel electrophoresis
Horizontal-gel electrophoresis
Vertical-gel electrophoresis

12. AIR DISPLACEMENT MICROPIPETTS
INVENTED BY: Dr.Heinrich Schnitger Marburg (1960)
Air displacement micropipettes are a type of adjustable
micropipette that deliver a measured volume of liquid; depending
on size, it could be between about o.1 ul to 1000 μl (1 ml). These
pipettes require disposable tips that come in contact with the
fluid.
PRINCIPLE :These pipettes operate by piston-driven air
displacement. A vacuum is generated by the vertical travel of a
metal or ceramic piston within an airtight sleeve. As the piston
moves upward, driven by the depression of the plunger, a
vacuum is created in the space left vacant by the piston. The
liquid around the tip moves into this vacuum (along with the air
in the tip) and can then be transported and released as necessary.
These pipettes are capable of being very precise and accurate.

14. ULTRA LOW TEMPERATURE FREEZERS
Invented by: Chuan Weng, Allan Kelly
• The instrument group ULT freezer is defined as freezers for
-80 to -85°C. ULT is the shortcut for ultra low temperature. There are
upright and chest freezers. The inner volume is in general between
300 and 800 L.
Principle: The refrigeration system of the ultra freezers basic cascade refrigeration principle,
the choice of two hermetic compressors as high, the compressor of the cryogenic stage. The
cryogenic stage system is also equipped with gas heat exchanger, allows low pressure gas from
the evaporator heat exchange with the high-pressure gas condensate evaporator, it will not only
reduce the heat load of the condensate evaporator, and the full use of the heat.
• Uses: for long term storage for biological samples like DNA, RNA, proteins, cell extracts, or
reagents. To reduce the risk of sample damage, these types of samples need extremely low
temperatures as -80 to -85°C.

15. INCUBATORS .
INVENTED BY: Louis pasteur
An incubator is a device used to grow and maintain
microbiological cultures or cell cultures. The incubator
maintains optimal temperature, humidity and other
conditions such as carbon di oxide and oxygen content of
atmosphere inside.
• PRINCIPLE:
An incubator has a compressor that works as a heater as well
as cooler and maintains the optimum or required temperature
for growth.

16. WEIGHT BALANCE
• A weighing balance is an instrument which is used to determine
the weight or mass of an object.
• Analytical balances are sensitive and expensive instruments, and
upon their accuracy and precision the accuracy of analysis result
depends.
• The most widely used type of analytical balances is balances with
a capacity of 100 g and a sensitivity of 0.1 mg. Not one
quantitative chemical analysis is possible without usage of
balances, because, regardless of which analytical method is being
used, there is always a need for weighing a sample for analysis
and the necessary quantity of reagents for solution preparation.
• The principle of operation of a modern laboratory balance bears some resemblance to its
predecessor the equal arm balance. The older instrument opposed the torque exerted by an
unknown mass on one side of a pivot to that of an adjustable known weight on the other side.
When the pointer returned to the center position, the torques must be equal, and the weight was
determined by the position of the moving weights.

17. • Types of weight Balance
a. Analytical Balance: 100mg-10g
b. Micro Balance : 2.0 mg-20.0 , Semi Micro Balance: 5ug- 500mg
c. Top load Balance : 1.0 g- 2.0g
• Modern electronic laboratory balances work on the
principle of magnetic force restoration. In this
system, the force exerted by the object being weighed
is lifted by an electromagnet. A detector measures the
current required to oppose the downward motion of
the weight in the magnetic field.
Analytical weight Balance
Micro Balance
Semi Micro Balance
Top load Balance

18. VORTEX MIXER
Invented by the Kraft brothers (Jack A. Kraft and Harold D. Kraft)
vortex mixer, or vortexer, is a simple device used commonly in laboratories
to mix small vials of liquid. It consists of an electric motor with the drive
shaft oriented vertically and attached to a cupped rubber piece mounted
slightly off-center. As the motor runs the rubber piece oscillates rapidly in a
circular motion. When a test tube or other appropriate container is pressed
into the rubber cup (or touched to its edge) the motion is transmitted to the
liquid inside and a vortex is created.
.
Principle:
Vortex in fluid dynamics, a vortex is a region in a fluid in which the flow rotates around an axis
line, which may be straight or curved.

19. PESTLE AND MORTAR
• A mortar and pestle is a kitchen device used since ancient
times to prepare ingredients or substances by crushing and
grinding them into a fine paste or powder. The mortar is a
bowl, made of hardwood, ceramic or stone. The pestle is a
heavy blunt club shaped object, end of which is used for
crushing and grinding.
• Uses :
It is used for grinding plant samples which lead to disrupting
cellular membranes and specially cell wall. Or in other words
to release biological molecules from inside the cell.

20. EPPENDROFF TUBES
• Are small capped plastic tubes used for centrifuge or in pcr
apparatus.
• Available in different volumes like 0.5 ml, 1.5 ml, 2 ml but
the most common size is 1.5 ml.
GLOVES AND UV GLASSES OR FACE SHIELDS .
Laboratory gloves are made of latex and nitrile. They protect
the hands of wearer against chemicals which may be corrosive
or carcinogenic or hazrdous in any nature. Do not use vinyl
gloves which can transmit significant amount of uv.
• Uv glasses or face shields: use poly carbonate face shields
that are rated for uv protection. It should be marked with 287 to
indicate that the shield meets the standard.
EPPENDROFF TUBES
GLOVES
UV GLASS
FACE SHIELDS

21. Microscopy
The morphological study of bacteria requires the use of microscopes. Microscopy has come a long
way since Leeuwenhoek first observed bacteria using handground lenses.
The types of microscope are
(i) Light or optical microscope
(ii) Phase contrast microscope
(iii) Dark field/ Dark ground microscope
(iv) Electron microscope

22. Light or optical microscope
They are of two types namely Simple and Compund Microscope
• Simple Microscope consists of a single lens. A hand lens is an
example of a simple Microscope.
• Compound Microscope consists of two or more lenses in series.
The image formed by the first lens is further magnified by another
lens.
Bacteria may be examined under the compound microscope, either
in the living state or after fixation and staining. Examination of wet
films or hanging drops indicates the shape, arrangements, motility
and approximately size of the cells. But due to lack of contrast
details cannot be appreciated.
Compound Microscope

23. Phase contrast microscope
-This imposes the contrast and makes evident the
structure within the cells that differ in thickness or
refractive index. The difference in the refractive
index between bacteria cells and the surrounding
medium makes them clearly visible.
-Retardation, by a fraction of a wavelength, of the
rays of light that pass through the object, compared
to the rays passing through the surrounding medium,
produces phase difference between the two types of
rays.

24. Dark field / Dark ground microscope
-Another method of improving the contrast is
the dark field microscope in which reflected
light is used instead of the transmitted light
used in the ordinal microscope.
-The contrast gives an illusion of increased
resolution, so that very slender organisms such
as spirochete, not visible under ordinary
illumination, can be clearly seen under the
dark field microscope.

25. Electron Microscope
- Beams of electron are used instead of beam of light, used
in light microscope.
- The object which is held in the path of beam scatters the
electrons and produces an image which is focused on a
fluorescent viewing screen.
- Gas molecules scatter electron, therefore it is necessary to
examine the object in a vacuum.

26. AUTOCLAVING:
-By using the autoclave, in which items are sterilized by
exposure to steam at 121º c and 15lbs of pressure for 15
minutes (or 126º c and 20lbs for 3 minutes).
-Air has been expelled and only steam is present in the
autoclave chamber.
-All forms of microbial life will be destroyed by this
method.

28. Spectrophotometer
Invented by: Arnold O. Beckman in 1940
-A spectrophotometer is an analytical instrument used
to quantitatively measure the transmission or reflection of
visible light, UV light or infrared light.
-Principle is that each compound absorbs or transmits light
over a certain range of wavelength
Spectrophotometer can be classified into two different types :
1. SINGLE BEAM SPECTRMETER:
To measure the intensity of the incident light the sample must be removed so that the reference can
be placed each time. This type of spectrometer is usually less expensive and less complicated.
2. DOUBLE BEAM SPECTOMETER:
In this type, before it reaches the sample, the light source is split into two separate beams. From
these one passes through the sample and second one is used for reference. This gives an advantage
because the reference reading and sample reading can take place at the same time.

29. Based on the wavelength of light used it can be classified into:
• VISIBLE SPECTROMETER
Uses visible range (400-700nm) of electromagnetic radiation spectrum Visible
spectrophotometers vary in accuracy. Plastic and glass cuvettes can be used for visible light
spectroscopy.
• UV SPECTOMETER
Uses light over the UV range (180-400 nm). UV spectroscopy is used for fluids, and even
solids. Cuvettes, only made of quartz, are used for placing the samples.
• IR SPECTROPHOTOMETER
Uses light over infra red range (700-15000) of electromagnetic radiation spectra.
-The spectrometer consists of the following parts:
1. Light source: it produce a desired range of wavelength of light.
2. Collimator: transmits a straight beam of light.
3. Monochromator: split the light into its component wavelength.
4. Wavelength selector transmits only the desired wavelength.

30. LAMINAR AIR FLOW CHAMBER
INVENTED BY: Willis Whitfield
• A laminar flow chamber is designed to prevent
contamination of semiconductor wafers, biological
samples, or any particle sensitive materials .
• Air is passed through a HEPA (High Efficiency
Particulates Air) filter which removes all airborne
contamination to maintain sterile conditions.
• Laminar air flow systems are used in various applications
such as life science research, mushroom cultivation,
microbiology, IVF, IUI and histopathology / pathology
lab, plant tissue and cell culture and pharmaceutical and
electronics industry.
• In the laboratory, Laminar Flow Cabinets are commonly
used for specialized work.

31. Parts of Laminar Air Flow
• A laminar flow hood consists of a filter pad, a fan and a HEPA (High Efficiency
Particulates Air) filter
• The fan sucks the air through the filter pad where dust is trapped .
• After that the prefiltered air has to pass the HEPA filter where contaminating fungi,
bacteria, dust etc are removed
• Sterile air flows into the working (flasking) area where you can do all your flasking
work without risk of contamination.
Types of Laminar Flow Cabinets
- Laminar Flow Cabinets can be produced as both horizontal and vertical cabinets
- There are many different types of cabinets with a variety of airflow patterns for different
purpose.
• Vertical Laminar Flow Cabinets
• Horizontal Laminar Flow Cabinets
• Laminar Flow Cabinets and Hoods
• Laminar Flow Benches and Booths

33. THANKYOU