safety and guidelines in molecular biology laboratory

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



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.



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.



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.



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.



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.



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),



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.



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

safety and guidelines in molecular biology laboratory

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