safety and guidelines in molecular biology laboratory
TRANSCRIPT
Molecular Biotechnology-Practic Lab-1 4th Grade Medical analysis February 11, 2020
1 Practical Molecular Biotechnology Muhsin Jamil Abdulwahid Lecturer in Molecular Genetics
Safety and Guidelines in molecular biology laboratory
Introduction Working in any kind of laboratory will require the analyst to possess a basic set of
practical skills. Measuring mass or volume, and preparing solutions of known
concentration, are likely to be everyday activities. In addition, working in a molecular
biology laboratory will entail specific considerations related to the nature of the
materials being handled.
Samples analysed in molecular biology laboratories are frequently of biological nature
– human or animal tissue − which can pose health risks to the analyst. Frequently, the
analyte of interest is DNA or RNA which, with current technologies, will need to be
amplified in order to be detected. This amplification step is susceptible to cross-
contamination from previously amplified material. Therefore, preventing
contamination of the samples and protecting the analyst are critical concerns for
molecular biologists.
Working in the laboratory The laboratory is a potentially hazardous working environment. You may well be
using chemicals, biological materials and equipment which, if not handled correctly,
could cause you or your colleagues harm. However, with the proper procedures in
place, work can be carried out safely. It is essential that you familiarise yourself with
the general safety procedures in place in your laboratory and with any special
procedures required to carry out a particular test method safely. When working in the
laboratory you also need to know how to select an appropriate test method and
equipment, and understand the importance of following standard operating procedures.
General Safety Rules and Procedures:
You should know the meaning of common warning and hazard signs used in the
laboratory. You will see different coloured signs:
Blue signs are mandatory(involuntary)
Red signs are prohibitive (or relate to fire alarms/fire-fighting equipment)
Green signs give safety instructions
Yellow signs give warnings
Molecular Biotechnology-Practic Lab-1 4th Grade Medical analysis February 11, 2020
2 Practical Molecular Biotechnology Muhsin Jamil Abdulwahid Lecturer in Molecular Genetics
Lab safety rules
1. No food or drinks are permitted in the laboratory at any time. 2. Keep hands and other objects away from your face, nose, eyes, ears, and mouth.
3. The application of cosmetics is prohibited in the laboratory.
4. Work areas / surfaces must be disinfected before and after use.
5. Laboratory coats must be worn.
6. Protective eyewear must be worn when performing any exercise or procedure in
the laboratory.
7. Long hair should be secured behind your head.
8. Hands must be washed before leaving the laboratory.
9. Never pipette anything by mouth (including water), always use pipetting
devices.
10. Label all materials with your name, date and any other applicable
information.
11. Dispose of wastes in their proper containers.
12. When handling chemicals note the hazard code on the bottle and take the
appropriate precautions indicated.
13. Do not pour biohazard's fluids down the sink.
Molecular Biotechnology-Practic Lab-1 4th Grade Medical analysis February 11, 2020
3 Practical Molecular Biotechnology Muhsin Jamil Abdulwahid Lecturer in Molecular Genetics
14. Glassware should be washed with soap and water, and then rinsed
with distilled water.
15. Immediately, report any broken glass, especially those containing
infectious materials.
Molecular Biotechnology-Practic Lab-1 4th Grade Medical analysis February 11, 2020
4 Practical Molecular Biotechnology Muhsin Jamil Abdulwahid Lecturer in Molecular Genetics
Equipment in molecular biology Laboratory
Biosafety Cabinets
Biosafety Cabinets (BSCs) are enclosed
workspaces with a ventilated hood that is
designed to contain pathogenic
microorganisms during microbiological
processes.
Molecular Biotechnology-Practic Lab-1 4th Grade Medical analysis February 11, 2020
5 Practical Molecular Biotechnology Muhsin Jamil Abdulwahid Lecturer in Molecular Genetics
Micropipette
Pipettes are an essential laboratory tool used to transport and dispense measured
volumes of liquids. Pipettes that dispense between 1 and 1000µl are termed
micropipettes, while macropipettes dispense a greater volume. Pipettes enable
sterile and accurate liquid handling and are commonly used within molecular biology,
analytical chemistry and medical tests. Pipettes come in several designs including
manual and electronic, as well as single-channel or multi-channel pipettes, all
designed for efficiency when working with varying plates, tubes, and dishes.
Molecular Biotechnology-Practic Lab-1 4th Grade Medical analysis February 11, 2020
6 Practical Molecular Biotechnology Muhsin Jamil Abdulwahid Lecturer in Molecular Genetics
Pipette Tips
Pipette Tips are disposable, autoclavable attachments for
the uptake and dispensing of liquids using a micropipette.
The volume of microliters each tip can hold varies, 0.5-
10µL, 2-20µL, 20-200µL, and 200-1000µL (1mL) are the
four standard tips. Pipette Tips are made of polypropylene
and are clear to allow for easy view of the tips contents.
Micropipette tips can be purchased non-sterile or sterile
both of which should be DNase, RNase, DNA, and
pyrogen free this is especially important for molecular
assays.
Eppendorf Tubes Eppendorf Tubes are single-use tubes made from polypropylene
for preparing, mixing, centrifuging, transporting and
storing solid and liquid samples and reagents. The
product can be used for training, routine and research
laboratories in the areas of life sciences, industry or
chemistry.
Molecular Biotechnology-Practic Lab-1 4th Grade Medical analysis February 11, 2020
7 Practical Molecular Biotechnology Muhsin Jamil Abdulwahid Lecturer in Molecular Genetics
PCR Tube Strips and Cap Strips
PCR Tube Strips and Cap Strips are designed for single use in PCR applications. PCR
tubes are small tubes made of high-quality virgin polypropylene with a conical bottom
and snap-cap lead. They have uniform thin walls to facilitate efficient heat transfer to
the sample. These tubes are autoclavable and work well with most thermal cyclers
Thermocycler Machine
A very early PCR machine which, rather than cycling through different
temperatures, uses three different water baths at constant temperatures
between which samples are moved with a robotic arm
Conventional PCR:
The thermal cycler (also known as a thermocycler, PCR machine or
DNA amplifier) is a laboratory apparatus most commonly used to
amplify segments of DNA via the polymerase chain reaction (PCR)
Real-Time PCR
A real-time polymerase chain reaction (real-time PCR), also
known as quantitative Polymerase Chain Reaction (qPCR), is a
laboratory technique of molecular biology based on the
polymerase chain reaction (PCR). It monitors the amplification
of a targeted DNA molecule during the PCR (i.e., in real time),
Molecular Biotechnology-Practic Lab-1 4th Grade Medical analysis February 11, 2020
8 Practical Molecular Biotechnology Muhsin Jamil Abdulwahid Lecturer in Molecular Genetics
not at its end, as in conventional PCR. Real-time PCR can be used quantitatively
(quantitative real-time PCR).
Vortex mixer
A 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.
Microcentrifuge
A microcentrifuge, also called a microfuge, it is used to spin small
(2 ml or less) liquid samples at high speeds (generally tens of
thousands times g-force).
UV transilluminators
Gel doc, also known as a gel documentation system, gel
image system or gel imager are used in molecular biology
laboratories for imaging , documentation or to view DNA
or RNA that has been separated by electrophoresis through
an agarose gel , these gels are typically stained with
ethidium bromide. Exposing the stained gel to a UV light
source causes the DNA to fluoresce and become visible.
These instruments are useful for viewing samples to size a
PCR product, purify DNA segments after a restriction
enzyme digest, quantify DNA, or verify RNA integrity
after extraction.
Molecular Biotechnology-Practic Lab-1 4th Grade Medical analysis February 11, 2020
9 Practical Molecular Biotechnology Muhsin Jamil Abdulwahid Lecturer in Molecular Genetics
Gel electrophoresis apparatus
The gel electrophoresis apparatus consists of a gel, which is often made from agar or polyacrylamide, and an
electrophoretic chamber (typically a hard plastic box or tank) with a cathode (negative terminal) at one end
and an anode (positive terminal) at the opposite end. The gel, which contains a series of wells at the cathode
end, is placed inside the chamber and covered with a buffer solution. The samples are then loaded into the
wells with a pipette. The chamber is connected to a power supply that, when turned on, applies an electric
field to the buffer.
NanoDrop Spectrophotometer:-
This system allows scientists to quickly and easily quantify and assess purity of samples
such as proteins and nucleic acids nucleic acid concentrations in sample volumes of one
microliter. ... This is accomplished by placing the sample directly on top of the detection
surface and using the surface tension to create a column between the ends of optical fibers.
An illustration of gel electrophoresis for DNA, showing the gel and
electrophoretic apparatus (left) and the separated bands of dyed