unisel

UNIVERSITI SELANGOR

FACULTY OF SCIENCE AND
BIOTECHNOLOGY
(FasBIO)
MOLECULAR TECHNIQUES BBS 1231
(LECTURER: MDM. Yasotha a/p Sundaraj)

GROUP 3
Devaraj a/l Ravindran 4111018161
Jeevitha a/p Tana sakaran 4111016861
Sujatha a/p kanniappan 4111009531
Premkumar a/l Subramaniam 4111017391
Theevindran a/l Kesavan 4111017311
Paveanthen a/l Ramachandran 4111023071


INTRODUCTION:
Molecular techniques are important skills to be mastered in order to perform well in areas
of biology, chemistry and genetics. Some of the molecular techniques are gel
electrophoresis, polymerase chain reaction, DNA scanning and imaging. Since these
techniques are advanced in terms of technology used for them, thus the equipments used
to carry out these techniques are also advance and complex too. The sophisticated
equipments must be handled in a proper manner in order to make a full use of it. Other
than that, equipments such as ph meter, multi channel pipette and electron microscope
must be calibrated according to their specified timing to ensure their precision, consistency
and accuracy.

AIM:
To identify the common equipments used by molecular biologist and understand their
functions and their ways of handling.

EQUIPMENTS:
Centrifuge, freezer, gel electrophoresis chamber, light microscope, microwave, pH meter,
thermocycler, uv transilluminator, water bath, incubator, shaker, weighing balance, electron
microscope, multi channel pipette, HPLC unit- high performance liquid chromatography.
METHOD:
Take a tour around the molecular biology laboratory in the university to identify common
equipments in the lab and listen to explanation given by the lecturer on those equipments.
Besides that, more equipment should be identified and studied by referring to other sources
such as text books and internet.

COMMON INSTRUMENTS IDENTIFIED AND STUDIED:
CENTRIFUGE
Centrifuge is an apparatus used to separate components in solution based on their density.
This machine has a metal block with holes to hold centrifuge tube and spins at desired
speed (rpm) and also for desired period of time. Upon rotation of centrifuge tubes around a
fixed axis, the components in the solution sediments according to their densities. Denser
substance sediments at the bottom of the tube meanwhile lighter substance tend to reside
on top of the denser substance. The capacity of the centrifuge tube ranges from 1.5ml to
15ml or even larger than that depending on the specification of the machine. Bench top
centrifuge machine tend to have smaller capacity of centrifuge tube. Centrifuge machine
also comes in smaller unit called mini centrifuge. Mini centrifuge is portable, used for short
period of time and quick spin.



Centrifuge (brand: ependorf) centrifuge tubes

FREEZER
Freezers are used to store samples at a very low temperature and for a long term. Normal
freezers tend to store samples at about -20 degrees Celsius thus those samples could be
preserved for a couple of months. Meanwhile, deep freezers are able to store samples at
about -80 degrees Celsius and those samples can be preserved for about one year.

Freezer deep freezer


GEL ELECTROPHORESIS
(electrophoresis chamber)
Gel electrophoresis is a method of using gel as a medium to sort out molecules such as
deoxyribonucleic acid(DNA) based on size and charge. This is made possible by the
movement of charged particle in an electrical field. The gel placed in an electrophoresis
chamber is connected to power source, when electric current is applied, the larger
molecules move slowly through the gel than smaller molecules. Then ,distinct bands are
formed. In order to make those bands to be visible, staining agent is added to the gel. A
photograph is also taken under ultraviolet light which makes the bands appear brighter.
(bands of DNA)

LIGHT MICROSCOPE
Light microscope is used to observe tiny objects that are not visible for naked eye. This
device has one ocular lens and three objective lens with various magnification powers (x10,
x40, x100). It also has coarse and fine adjustment knobs to focus on specimen placed on the
stage. The lights source to observe the specimen comes from a light source under the
diaphragm. In some less advanced microscope, the light source comes from the reflection
of light on the mirror placed under the diaphragm.


MIRCOWAVE OVEN
Microwave oven heats up substance placed in it by passing through non ionizing microwave
radiation. The heating compartment of the oven is made up of aluminium and consists of
transmitters to transmit microwaves. The heating temperature and timing can be set
manually before using it. In laboratory, microwave ovens are used to prepare agarose gel
for DNA transfer.
(1000 watt microwave oven)




PH METER
pH meter is an electronic instrument used for measuring the pH of a liquid. A laboratory pH
meter consists of a special measuring probe connected to an electronic meter that
measures and displays the pH reading. pH value is measured based on hydrogen cations in
the solution. When the probe is dipped into a solution, the hydrogen ions in it surround the
thin-walled glass bulb (probe). The activity of the hydrogen ions causes the probe to
produce a small voltage and it will be measured and displayed as pH units by the meter. The
meter is similar to voltmeter but the measurements are displayed in pH units instead of
volts.
The pH meter has to be calibrated before using because the glass electrode does not give a
reproducible electromotive force over longer periods of time. Calibration is performed with
at least two standard buffer solutions. The glass probe of the pH meter is the most crucial
and sensitive part. It must be kept wet at all times to avoid dehydration of pH sensing
membrane and dysfunction of electrode. . After using the pH meter for every measurement,
the probe is rinsed with distilled water to remove any traces of the tested solution.

Labrotary pH meter portable pH meter







THERMOCYCLER PCR MACHINE
Thermocycler PCR machine or DNA amplifier is a machine or laboratory apparatus that uses
Polymerase chain reaction process to amplify segments of DNA. This machine is very useful
when a limited amount of DNA sample need to go through various tests. A thermocycler
PCR machine can produce thousands or millions of copies of an original sample to be tested.
This machine comes in various designs from various biotech companies, however the
function and working principles are the same. This machine has a termal block with holes to
insert tubes containing PCR reaction mixtures. The machine then changes the temperature
of the block in pre-programmed steps.


thermocyclers of three different brands





UV TRANSILLUMINATOR and CELL DOCUMENTATON SYSTEM
UV transilluminator is a device used to observe bands especially DNA bands found in the
agarose gel after being electrophoresised. Some components in the agarose gel fluoresce
under UV light, thus the gel will be placed on an UV transilluminator for visualization
purpose. This equipment comes in two forms: with and without darkbox design. Darkbox
design illuminator has a digital camera to capture the image and save it in a PC unit
connected to it. The image captured can be even printed out for other purposes.

UV transilluminator with darkbox

UV transilluminator without darkbox
LABORATORY WATER BATH
Laboratory water bath is used to maintain a very stable temperature around any vessels
containing reacting substances or chemicals, therefore it can be also used for incubation
process. This is made possible by heating and circulating the water in the tank. Other than
that, this equipment can also be used to heat substances that cannot be heated by Bunsen
burner. The temperature of the water can be set before using it. Water baths can hold
temperatures within 100 degrees Celsius.


INCUBATOR
An incubator is a device used to grow and maintain microbiological cultures or cell cultures.
The incubator maintains optimal temperature and humidity of the atmosphere inside.
Incubators are essential for a lot of experimental work in molecular biology and are used to
culture both bacterial as well as eukaryotic cells. Most incubators include a timer which can
be programmed to cycle through different temperatures and humidity levels. Incubators can
vary in size from tabletop to units the size of small rooms. The simplest incubators are
insulated boxes with an adjustable heater, typically going up to 60 to 65 C. Commonly in
microbiological cultures the temperature that is used frequently is 37 C for the organisms
that grows under these conditions (bacteria). Other biological organisms such as yeast the
beat temperature used to culture it are 30 C.
There are many different types and sizes of incubators. Uni-therm incubators, mini
incubators, refrigerated incubators, humidity and constant temperature incubators, uni-
therm carbon dioxide incubator, unihood incubation hood and cooled incubators are the
ones which are mostly used in many laboratory experiments










Uni-therm Carbon Dioxide Incubator
Uni-therm Incubator
Uni-therm Refrigerated Incubator
Uni-hood Incubation Hood


SHAKER

SHAKER
Shakers are used to culture microorganisms and algae in molecular biology. They shake and
vibrate the microorganisms in the beaker that are placed on the shaker. The main purpose
of doing this is to stir the solution of the microorganism. The units that are used in shakers
are rpm which stands for round per minute. Shakers are mainly powered by an electric
motor. Shakers are classified into two which are heated shaker and unheated shaker. Both
this shaker owns different characteristics but with same function. It has indexing device and
timer. The frequency of this unheated shaker is from 40 to 200 rpm with the amplitude of
20mm. Another unheated shaker has horizontal shaking motion. It also has indexing device
and timer as the other one. The frequency of this type is 50 to 1100 rpm with amplitude of
3mm.
Heated shaker are the shakes with heated platform. Its a microprocessor controlled thermo
shaker for 4 micro liter plates with -65C RT and 3mm orbit. Heated shaker, shakes
frequently throughout the particular period of time.




Humidity Incubator Mini Incubator
Shaker
WEIGHING BALANCE
Weighing balance is used to measure mass to a very high degree of precision and accuracy.
The measuring pan of a high precision weighing balance are placed inside a transparent
enclosure with doors to avoid dust from being collect and from any air currents in the room
do not affect the balance's operation. The use of a vented balance safety enclosure, which
has uniquely designed acrylic airfoils, allows a smooth turbulence-free airflow that prevents
balance fluctuation and the measure of mass down to 1 g without fluctuations or loss of
product. Also, the sample must be at room temperature to prevent natural convection from
forming air currents inside the enclosure, affecting the measure of mass.
Weighing precision is achieved by maintaining a constant load on the balance beam, by
subtracting mass on the same side of the beam to which the sample is added. The final
balance is achieved by using a small spring force rather than subtracting fixed masses.











Weighing Balance
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY UNIT HPLC UNIT
HPLC unit is a tool used to carry out a highly improved liquid chromatography. In a normal
liquid chromatography tool, a solvent is allowed to drip through a column under the
influence of gravity but in a HPLC, solvent is forced through under high pressures up to 400
atmospheres which makes it much faster. The components of this HPLC unit are solvent
reservoir, a pump to produce high pressure, HPLC tube, detector and a processing unit to
display it. This equipment allows a better separation of the components of the mixture. The
injection of sample into the HPLC tube is an automated process.

HPLC unit

CONCLUSION:
Common equipments used by molecular biologist, their function and the ways to handle
them are identified and learned.

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